Comparison of Metal Nanoparticles (Au, Ag, Eu, Cd) Used for Immunoanalysis Using LA-ICP-MS Detection

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Authors

VLČNOVSKÁ Marcela ŠTOSSOVÁ Aneta KUCHYNKA Michaela FALTUSOVÁ Veronika HOLCOVÁ POLANSKÁ Hana MASAŘÍK Michal HRSTKA Roman ADAM Vojtech KANICKÝ Viktor VACULOVIČ Tomáš VACULOVIČOVÁ Markéta

Year of publication 2021
Type Article in Periodical
Magazine / Source Molecules
MU Faculty or unit

Faculty of Medicine

Citation
Web https://www.mdpi.com/1420-3049/26/3/630
Doi http://dx.doi.org/10.3390/molecules26030630
Keywords protein p53; dot-blot; antibody
Description Immunochemical methods are used not only in clinical practice for the diagnosis of a wide range of diseases but also in basic and advanced research. Based on the unique reaction between the antibody and its respective antigens, it serves to specifically recognize target molecules in biological complex samples. Current methods of labelling antibodies with elemental labels followed by detection by inductively coupled plasma mass spectrometry (ICP-MS) allow detection of multiple antigens in parallel in a single analysis. Using the laser ablation (LA) modality (LA-ICP-MS), it is also possible to monitor the spatial distribution of biogenic elements. Moreover, the employment of metal nanoparticle-labeled antibodies expands the applicability also to molecular imaging by LA-ICP-MS. In this work, conjugates of model monoclonal antibody (DO-1, recognizing p53 protein) with various metal nanoparticles-based labels were created and utilized in dot-blot analysis in order to compare their benefits and disadvantages. Based on experiments with the p53 protein standard, commercial kits of gold nanoparticles proved to be the most suitable for the preparation of conjugates. The LA-ICP-MS demonstrated very good repeatability, wide linear dynamic range (0.1-14 ng), and limit of detection was calculated as a 1.3 pg of p53 protein.
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