Ternary covalent conjugate (antibody-gold nanoparticle-peroxidase) for signal enhancement in enzyme immunoassayстатья
Статья опубликована в высокорейтинговом журнале
Информация о цитировании статьи получена из
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Статья опубликована в журнале из списка Web of Science и/или Scopus
Дата последнего поиска статьи во внешних источниках: 1 августа 2016 г.
Аннотация:A synthesis of new conjugate of horseradish peroxidase (HRP) and antibody covalently bound with gold nanoparticles (GNPs) with a high content of the enzyme was developed first. Varying the concentrations of the reacting compounds the synthesis conditions were optimized. In the conjugate, which was synthesized under the optimized conditions, one gold nanoparticle carrier (Ø 30 nm) at its surface contains 13 molecules of active HRP and 1 molecule of anti-troponin I antibody. The applicability of the proposed ternary conjugate was successfully proved in chemiluminescent ELISA for the determination of human troponin I. The sensitivity of the assay using the ternary conjugate was higher than that of the assay with the streptavidin-biotin system (biotinylated antibody and streptavidin-HRP conjugate). High sensitivity was reached due to the dual signal amplification from use of a potent enhancer of HRP-catalyzed chemiluminescence (3-(10′-phenothiazinyl)propane-1-sulfonate) and the high content of HRP in the conjugate. A replacement of two-steps streptavidin-biotin amplification system with the ternary conjugate decreased the time of the assay. The use of the proposed conjugate allowed significantly reducing a consumption of antibody and HRP per assay.