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@article{2417981,
author = {Máčala, Jakub and Makhneva, Ekaterina and Hlaváček, Antonín and Kopecký, Martin and Gorris, HansandHeiner and Skládal, Petr and Farka, Zdeněk},
article_location = {Washington, DC},
article_number = {25},
doi = {http://dx.doi.org/10.1021/acs.analchem.4c00837},
keywords = {Photon-upconversion nanoparticle; Immunoassay; Dot-blot; Biomarker; Human serum albumin; Prostate-specific antigen; Cardiac troponin},
language = {eng},
issn = {0003-2700},
journal = {Analytical Chemistry},
title = {Upconversion Nanoparticle-Based Dot-Blot Immunoassay for Quantitative Biomarker Detection},
url = {https://pubs.acs.org/doi/10.1021/acs.analchem.4c00837},
volume = {96},
year = {2024}
}
TY - JOUR
ID - 2417981
AU - Máčala, Jakub - Makhneva, Ekaterina - Hlaváček, Antonín - Kopecký, Martin - Gorris, Hans-Heiner - Skládal, Petr - Farka, Zdeněk
PY - 2024
TI - Upconversion Nanoparticle-Based Dot-Blot Immunoassay for Quantitative Biomarker Detection
JF - Analytical Chemistry
VL - 96
IS - 25
SP - 10237-10245
EP - 10237-10245
PB - American Chemical Society
SN - 00032700
KW - Photon-upconversion nanoparticle
KW - Immunoassay
KW - Dot-blot
KW - Biomarker
KW - Human serum albumin
KW - Prostate-specific antigen
KW - Cardiac troponin
UR - https://pubs.acs.org/doi/10.1021/acs.analchem.4c00837
N2 - Dot-blot immunoassays are widely used for the user-friendly detection of clinical biomarkers. However, the majority of dot-blot assays have only limited sensitivity and are only used for qualitative or semiquantitative analysis. To overcome this limitation, we have employed labels based on photon-upconversion nanoparticles (UCNPs) that exhibit anti-Stokes luminescence and can be detected without optical background interference. First, the dot-blot immunoassay on a nitrocellulose membrane was optimized for the quantitative analysis of human serum albumin (HSA), resulting in a limit of detection (LOD) of 0.19 ng/mL and a signal-to-background ratio (S/B) of 722. Commercial quantum dots were used as a reference label, reaching the LOD of 4.32 ng/mL and the S/B of 3, clearly indicating the advantages of UCNPs. In addition, the potential of UCNP-based dot-blot for real sample analysis was confirmed by analyzing spiked urine samples, reaching the LOD of 0.24 ng/mL and recovery rates from 79 to 123%. Furthermore, we demonstrated the versatility and robustness of the assay by adapting it to the detection of two other clinically relevant biomarkers, prostate-specific antigen (PSA) and cardiac troponin (cTn), reaching the LODs in spiked serum of 9.4 pg/mL and 0.62 ng/mL for PSA and cTn, respectively. Finally, clinical samples of patients examined for prostate cancer were analyzed, achieving a strong correlation with the reference electrochemiluminescence immunoassay (recovery rates from 89 to 117%). The achieved results demonstrate that UCNPs are highly sensitive labels that enable the development of dot-blot immunoassays for quantitative analysis of low-abundance biomarkers.
ER -
MÁČALA, Jakub, Ekaterina MAKHNEVA, Antonín HLAVÁČEK, Martin KOPECKÝ, Hans-Heiner GORRIS, Petr SKLÁDAL a Zdeněk FARKA. Upconversion Nanoparticle-Based Dot-Blot Immunoassay for Quantitative Biomarker Detection. \textit{Analytical Chemistry}. Washington, DC: American Chemical Society, 2024, roč.~96, č.~25, s.~10237-10245. ISSN~0003-2700. Dostupné z: https://dx.doi.org/10.1021/acs.analchem.4c00837.
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