Upconversion Nanoparticle-Based Dot-Blot Immunoassay for
Quantitative Biomarker Detection

J 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 (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

URL

Impakt faktor

Impact factor: 7.400 v roce 2022

Organizační jednotka

Přírodovědecká fakulta

DOI

http://dx.doi.org/10.1021/acs.analchem.4c00837

UT WoS

001247434700001

Klíčová slova anglicky

Photon-upconversion nanoparticle; Immunoassay; Dot-blot; Biomarker; Human serum albumin; Prostate-specific antigen; Cardiac troponin

Štítky

rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 12. 7. 2024 13:46, Mgr. Marie Šípková, DiS.

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)

Název: Modernizace České infrastruktury pro integrativní strukturní biologii (Akronym: UP CIISB)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Modernizace České infrastruktury pro integrativní strukturní biologii, PO 1 Posilování kapacit pro kvalitní výzkum

EF18_046/0015974, projekt VaV

Název: Modernizace České infrastruktury pro integrativní strukturní biologii

GA22-27580S, projekt VaV

Název: Laserová spektroskopie v imunostanovení a zobrazování s nanometalickými značkami (Akronym: IMMUNOLIBS)

Investor: Grantová agentura ČR, Laserová spektroskopie v imunostanovení a zobrazování s nanometalickými značkami

LM2023042, projekt VaV

Název: Česká infrastruktura pro integrativní strukturní biologii

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, CIISB - Česká infrastruktura pro integrativní strukturní biologii

Citovat

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. 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.

@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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