2024
Upconversion Nanoparticle-Based Dot-Blot Immunoassay for Quantitative Biomarker Detection
MÁČALA, Jakub, Ekaterina MAKHNEVA, Antonín HLAVÁČEK, Martin KOPECKÝ, Hans-Heiner GORRIS et. al.Základní údaje
Originální název
Upconversion Nanoparticle-Based Dot-Blot Immunoassay for Quantitative Biomarker Detection
Autoři
MÁČALA, Jakub ORCID (203 Česká republika, domácí), Ekaterina MAKHNEVA (643 Rusko, domácí), Antonín HLAVÁČEK (203 Česká republika), Martin KOPECKÝ (203 Česká republika, domácí), Hans-Heiner GORRIS (276 Německo, domácí), Petr SKLÁDAL (203 Česká republika, domácí) a Zdeněk FARKA (203 Česká republika, garant, domácí)
Vydání
Analytical Chemistry, Washington, DC, American Chemical Society, 2024, 0003-2700
Další údaje
Jazyk
angličtina
Typ výsledku
Článek v odborném periodiku
Obor
10406 Analytical chemistry
Stát vydavatele
Spojené státy
Utajení
není předmětem státního či obchodního tajemství
Odkazy
Impakt faktor
Impact factor: 7.400 v roce 2022
Organizační jednotka
Přírodovědecká fakulta
UT WoS
001247434700001
Klíčová slova anglicky
Photon-upconversion nanoparticle; Immunoassay; Dot-blot; Biomarker; Human serum albumin; Prostate-specific antigen; Cardiac troponin
Příznaky
Mezinárodní význam, Recenzováno
Změněno: 10. 2. 2025 08:36, prof. RNDr. Petr Skládal, CSc.
Anotace
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.
Návaznosti
| CZ.02.1.01/0.0/0.0/18_046/0015974, interní kód MU (Kód CEP: EF18_046/0015974) |
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| EF18_046/0015974, projekt VaV |
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| GA22-27580S, projekt VaV |
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| LM2023042, projekt VaV |
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| MUNI/A/1582/2023, interní kód MU |
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