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Nanoparticle-based immunochemical assays: Nano-update of classic analytical tools

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FARKA Zdeněk HLAVÁČEK Antonín MICKERT Matthias Jürgen GORRIS Hans-Heiner SKLÁDAL Petr

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Fakulta / Pracoviště MU

Středoevropský technologický institut

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Popis Enzyme immunoassays are widely used for detection of various analytes within complex samples. However, enzymes as labels suffer several disadvantages such as high production costs, limited stability, and time-consuming signal development. The recent progress in nanotechnology has led to the development of various nanomaterials that can be used to overcome these disadvantages and improve the assay properties. Prussian blue nanoparticles (PBNPs) are nanozymes composed of the Fe4[Fe(CN)6]3-based coordination polymer. They reveal peroxidase-like activity and are capable of catalyzing the oxidation of tetramethylbenzidine to form intensely blue product. We have introduced the method for conjugation of PBNPs with antibodies and demonstrated the universal applicability of the label by the development of sandwich nanozyme-linked immunosorbent assays (NLISAs) for human serum albumin in urine and Salmonella Typhimurirum in powdered milk. Photon-upconversion nanoparticles (UCNPs) are lanthanide-doped nanocrystals, which convert near-infrared (NIR) light to light of shorter wavelengths (anti-Stokes emission). We have developed a competitive upconversion-linked immunosorbent assay (ULISA) for the detection of pharmaceutical diclofenac, which is a common micropollutant in waters. The low background and high photostability make UCNPs a powerful tool for detection of single molecules. We have developed an optical approach for visualizing individual UCNPs by epiluminescence microscopy and applied it for the sensitive detection of the cancer biomarker prostate specific antigen (PSA). The noise-surpassing digital readout provided single particle resolution with almost no instrumental background. The concept of introducing nanomaterials into immunoassays modernizes the classic bioanalytical methods and improves their working parameters. The digital signal readout is a completely new approach with the potential to pave the way for a new generation of immunoassays.
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