2024
Upconversion Nanoparticle-Based Dot-Blot Immunoassay for Quantitative Biomarker Detection
MÁČALA, Jakub, Ekaterina MAKHNEVA, Antonín HLAVÁČEK, Martin KOPECKÝ, Hans-Heiner GORRIS et. al.Basic information
Original name
Upconversion Nanoparticle-Based Dot-Blot Immunoassay for Quantitative Biomarker Detection
Authors
MÁČALA, Jakub ORCID (203 Czech Republic, belonging to the institution), Ekaterina MAKHNEVA (643 Russian Federation, belonging to the institution), Antonín HLAVÁČEK (203 Czech Republic), Martin KOPECKÝ (203 Czech Republic, belonging to the institution), Hans-Heiner GORRIS (276 Germany, belonging to the institution), Petr SKLÁDAL (203 Czech Republic, belonging to the institution) and Zdeněk FARKA (203 Czech Republic, guarantor, belonging to the institution)
Edition
Analytical Chemistry, Washington, DC, American Chemical Society, 2024, 0003-2700
Other information
Language
English
Type of outcome
Article in a journal
Field of Study
10406 Analytical chemistry
Country of publisher
United States of America
Confidentiality degree
is not subject to a state or trade secret
References:
Impact factor
Impact factor: 6.800 in 2023
Organization unit
Faculty of Science
UT WoS
001247434700001
Keywords in English
Photon-upconversion nanoparticle; Immunoassay; Dot-blot; Biomarker; Human serum albumin; Prostate-specific antigen; Cardiac troponin
Tags
International impact, Reviewed
Changed: 16/3/2025 16:11, Mgr. Eva Dubská
Abstract
V originále
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.
Links
CZ.02.1.01/0.0/0.0/18_046/0015974, interní kód MU (CEP code: EF18_046/0015974) |
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EF18_046/0015974, research and development project |
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GA22-27580S, research and development project |
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MUNI/A/1582/2023, interní kód MU |
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90242, large research infrastructures |
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