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.

Basic information

Original name

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

Authors

MÁČALA, Jakub (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

Článek v odborném periodiku

Field of Study

10406 Analytical chemistry

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 7.400 in 2022

Organization unit

Faculty of Science

DOI

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

UT WoS

001247434700001

Keywords in English

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

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)

Name: Modernizace České infrastruktury pro integrativní strukturní biologii (Acronym: UP CIISB)

Investor: Ministry of Education, Youth and Sports of the CR, Modernization of the Czech Infrastructure for Integrative Structural Biology, Priority axis 1: Strengthening capacities for high-quality research

EF18_046/0015974, research and development project

Name: Modernizace České infrastruktury pro integrativní strukturní biologii

GA22-27580S, research and development project

Name: Laserová spektroskopie v imunostanovení a zobrazování s nanometalickými značkami (Acronym: IMMUNOLIBS)

Investor: Czech Science Foundation

LM2023042, research and development project

Name: Česká infrastruktura pro integrativní strukturní biologii

Investor: Ministry of Education, Youth and Sports of the CR, CIISB - Czech Infrastructure for Integrative Structural Biology

Citovat

MÁČALA, Jakub, Ekaterina MAKHNEVA, Antonín HLAVÁČEK, Martin KOPECKÝ, Hans-Heiner GORRIS, Petr SKLÁDAL and Zdeněk FARKA. Upconversion Nanoparticle-Based Dot-Blot Immunoassay for Quantitative Biomarker Detection. Analytical Chemistry. Washington, DC: American Chemical Society, 2024, vol. 96, No 25, p. 10237-10245. ISSN 0003-2700. Available from: 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 and Zdeněk FARKA. Upconversion Nanoparticle-Based Dot-Blot Immunoassay for Quantitative Biomarker Detection. \textit{Analytical Chemistry}. Washington, DC: American Chemical Society, 2024, vol.~96, No~25, p.~10237-10245. ISSN~0003-2700. Available from: https://dx.doi.org/10.1021/acs.analchem.4c00837.

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