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Instantaneous determination via bimolecular recognition: usefulness of FRET in phosphorylcholine group enriched nanoparticles
Authors:Watanabe Junji  Ishihara Kazuhiko
Affiliation:Department of Applied Chemistry and 21st Century COE Program, Center for Integrated Cell and Tissue Regulation, Graduate School of Engineering, Osaka University, Suita, Osaka, Japan.
Abstract:This paper deals with smart bimolecular recognition for instantaneous determination. In particular, we installed the fluorescence resonance energy transfer (FRET) system in phosphorylcholine (PC) group enriched nanoparticles (NPs). The most favorable characteristics were as follows: (i) the suppression of nonspecific protein adsorption by the PC group enriched surface and (ii) simple bioassay protocol relative to the conventional enzyme-linked immunosorbent assay (ELISA). In the case of immunoassays, nonspecific interaction and complex protocols are known dominant problems. To address these issues, we designed FRET-installed NPs. Agglutination of NPs is a fundamental immunoassay technique; however, it is not quantitative. By evaluating the degree of agglutination based on the fluorescence intensity, the resulting information can be used for diagnosis. Therefore, we installed the FRET system on the surface of the NPs. In this paper, C-reactive protein (CRP) and osteopontin (OPN) were the target biomarkers for instantaneous determination, and the resulting fluorescence intensity correlated well with changes in the concentrations of the target molecules. The immunoassay protocol was quite simple, involving only the mixing of FRET-installed NPs and target molecules, such as CRP and OPN antigens. We successfully evaluated the concentration of the target biomarkers, even when human serum albumin was present as an interference molecule.
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