Facile Encapsulation of Iridium(III) Complexes in Apoferritin Nanocages as Promising Electrochemiluminescence Nanodots for Immunoassays.

A class of water-soluble electrochemiluminescence (ECL) nanodots were prepared by encapsulating ECL-active iridium complexes into biocompatible horse spleen apoferritin (apoHSF) nanocages for immunoassays. The preparation feasibility was achieved based on the pH-induced disassembly/reassembly nature originated from apoHSF. Two iridium nanodots (1 and 2) with high ECL efficiency were separately prepared by directing the self-assembly of two water-insoluble luminescent complexes, Ir(ppy)3 (ppy = 2-phenylpyridine) and Ir(ppy)2(acac) (ppy = 2-phenylpyridine and acac = acetylacetonate), in the apoHSF cavity. Using tri-n-propylamine (TPrA) as a coreactant, the electrochemistry and "oxidative-reductive" ECL mechanisms for nanodots 1 and 2 were investigated, respectively. After demonstrating the spectroscopic property and relative ECL efficiency, the ECL emission of nanodots 1 and 2 quenched by TPrA• radicals at high potential was further studied and circumvented by optimizing the potential range and TPrA concentration for generating strong and stable ECL emission in aqueous media. The well-inherited biological functions of apoHSF in nanodots allow the convenient external modification of an antibody to act as a signal probe, thus a versatile ECL immunoassay paradigm was established. Acceptable results from this assay enabled the rapid and accurate detection of biomarkers in real samples. The unprecedented use of apoHSF is feasible and applicable for water-insoluble iridium complexes to fabricate a wide variety of biocompatible ECL nanodots for potential bioanalysis.