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Prussian blue nanoparticles as nanozyme label in immunoassays

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ČUNDERLOVÁ Veronika HLAVÁČEK Antonín FARKA Zdeněk HORÁČKOVÁ Veronika MIKUŠOVÁ Zuzana SKLÁDAL Petr

Rok publikování 2018
Druh Konferenční abstrakty
Fakulta / Pracoviště MU

Středoevropský technologický institut

Citace
Popis Enzymes represent the most often used labels for amplification of signal in immunoassays due to their high catalytic efficiency. However, natural enzymes suffer from several disadvantages, e.g. they are easily denatured and their production and purification is generally complicated and time-consuming. Contrary, inorganic catalytic nanoparti-cles (nanozymes) offer easy and fast synthesis from commonly used chemicals. One of them are Prussian blue nanoparticles (PBNPs), which are nanozymes composed of coordination polymer of Fe4[Fe(CN)6]3. PBNPs contain the Fe3+/Fe2+ redox couple on the surface, which is also responsible for catalytic activity of peroxidases. PBNPs show high efficiency in catalyzing the oxidation of colorless 3,3,5,5-tetramethylbenzi-dine in the presence of H2O2 to form intensively blue product. We have prepared PBNPs with turnover number of 20 000 s-1 towards TMB oxidation, which is the highest value reported for nanoparticles of a size comparable to common proteins. We have also introduced the method for conjugation of PBNPs with antibody and utilization of the conjugate as a catalytic label in sandwich nanozyme-linked immunosorbent assay (NLISA). The practical applicability of NLISA was demonstrated by detection of different antigens, human serum albumin in urine and Salmonella Typhimurium in milk with limit of detection of 1.2 ng mL1 and 6·10^3 CFU mL1 respectively.
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