Abstract
The kosmotropic effect induced by ammonium sulfate (AS) at concentrations greater than approximately 2.8 M allows the marked intensification of chemiluminescence (CL) arising from a conventional luminol–hydrogen peroxide (H2O2)–horseradish peroxidase (HRP) reaction. Because of the kosmotropic effect, CL is intensified by at least three orders of magnitude than that from the conventional HRP-catalyzed luminol reaction with no AS; the linear relationship between the CL intensity and the HRP concentration is established over the range of 0.3 pM to several tens of pM. The novel CL intensification effect on the HRP-catalyzed luminol CL can be stably and reproducibly induced.Graphical abstract
Highlights
Chemiluminescence (CL), as a result of the chemiexcitation derived from a series of chemical reactions, is an effective tool in analytical chemistry [1]
In the enzyme-linked immunosorbent assay (ELISA), horseradish peroxidase (HRP) is used as a marker enzyme for the secondary antibodies and the highly sensitive detection of HRP-linked antibodies is attained based on the luminol CL [2]
The reaction mechanism for the CL arising from the luminol–H2O2–HRP reaction is described in Scheme S1 (Supporting Information)
Summary
Chemiluminescence (CL), as a result of the chemiexcitation derived from a series of chemical reactions, is an effective tool in analytical chemistry [1]. It was shown that the CL from an HRPcatalyzed luminol reaction was markedly intensified in the presence of high concentrations of AS by at least three orders of magnitude than that from the conventional luminol–H2O2–HRP reaction without using the so-called enhancer agent. The absorption spectra of various concentrations of HRP in the presence and absence of 3.5 M AS were measured, and the results are shown in Fig. S2 (Supporting Information).
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