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@article{1836099, author = {Hlaváček, Antonín and Farka, Zdeněk and Mickert, Matthias Jürgen and Kostiv, Uliana and Brandmeier, Julian and Horák, Daniel and Skládal, Petr and Foret, František and Gorris, HansandHeiner}, article_number = {April 2022}, doi = {http://dx.doi.org/10.1038/s41596-021-00670-7}, keywords = {Photon-upconversion nanoparticle; Immunoassay; Immunocytochemistry; Cancer biomarker; Bioconjugation}, language = {eng}, issn = {1754-2189}, journal = {Nature Protocols}, title = {Bioconjugates of photon-upconversion nanoparticles for cancer biomarker detection and imaging}, url = {https://www.nature.com/articles/s41596-021-00670-7}, volume = {17}, year = {2022} }
TY - JOUR ID - 1836099 AU - Hlaváček, Antonín - Farka, Zdeněk - Mickert, Matthias Jürgen - Kostiv, Uliana - Brandmeier, Julian - Horák, Daniel - Skládal, Petr - Foret, František - Gorris, Hans-Heiner PY - 2022 TI - Bioconjugates of photon-upconversion nanoparticles for cancer biomarker detection and imaging JF - Nature Protocols VL - 17 IS - April 2022 SP - 1028-1072 EP - 1028-1072 PB - Springer Nature SN - 17542189 KW - Photon-upconversion nanoparticle KW - Immunoassay KW - Immunocytochemistry KW - Cancer biomarker KW - Bioconjugation UR - https://www.nature.com/articles/s41596-021-00670-7 N2 - The detection of cancer biomarkers in histological samples and blood is of paramount importance for clinical diagnosis. Current methods are limited in terms of sensitivity, hindering early detection of disease. We have overcome the shortcomings of currently available staining and fluorescence labeling methods by taking an integrative approach to establish photon-upconversion nanoparticles (UCNP) as a powerful platform for cancer detection. These nanoparticles are readily synthesized in different sizes to yield efficient and tunable short-wavelength light emission under near-infrared excitation, which eliminates optical background interference of the specimen. Here we present a protocol for the synthesis of UCNPs by high-temperature co-precipitation or seed-mediated growth by thermal decomposition, surface modification by silica or poly(ethylene glycol) that renders the particles resistant to nonspecific binding, and the conjugation of streptavidin or antibodies for biological detection. To detect blood-based biomarkers, we present an upconversion-linked immunosorbent assay for the analog and digital detection of the cancer marker prostate-specific antigen. When applied to immunocytochemistry analysis, UCNPs enable the detection of the breast cancer marker human epidermal growth factor receptor 2 with a signal-to-background ratio 50-fold higher than conventional fluorescent labels. UCNP synthesis takes 4.5 d, the preparation of the antibody–silica–UCNP conjugate takes 3 d, the streptavidin–poly(ethylene glycol)–UCNP conjugate takes 2–3 weeks, upconversion-linked immunosorbent assay takes 2–4 d and immunocytochemistry takes 8–10 h. The procedures can be performed after standard laboratory training in nanomaterials research. ER -
HLAVÁČEK, Antonín, Zdeněk FARKA, Matthias Jürgen MICKERT, Uliana KOSTIV, Julian BRANDMEIER, Daniel HORÁK, Petr SKLÁDAL, František FORET a Hans-Heiner GORRIS. Bioconjugates of photon-upconversion nanoparticles for cancer biomarker detection and imaging. \textit{Nature Protocols}. Springer Nature, 2022, roč.~17, April 2022, s.~1028-1072. ISSN~1754-2189. Dostupné z: https://dx.doi.org/10.1038/s41596-021-00670-7.
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