Bioconjugates of photon-upconversion nanoparticles with
antibodies for the detection of prostate-specific antigen and
p53 in heterogeneous and homogeneous immunoassays

J 2025

Bioconjugates of photon-upconversion nanoparticles with antibodies for the detection of prostate-specific antigen and p53 in heterogeneous and homogeneous immunoassays

MAKHNEVA, Ekaterina; Pavel ŠPAČEK; Antonín HLAVÁČEK; Julie WEISOVÁ; Hans-Heiner GORRIS et al.

Základní údaje

Originální název

Bioconjugates of photon-upconversion nanoparticles with antibodies for the detection of prostate-specific antigen and p53 in heterogeneous and homogeneous immunoassays

Autoři

MAKHNEVA, Ekaterina; Pavel ŠPAČEK; Antonín HLAVÁČEK; Julie WEISOVÁ; Hans-Heiner GORRIS; Petr SKLÁDAL a Zdeněk FARKA

Vydání

Nanoscale, Royal Society of Chemistry, 2025, 2040-3364

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10406 Analytical chemistry

Stát vydavatele

Velká Británie a Severní Irsko

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 5.100 v roce 2024

Označené pro přenos do RIV

Ano

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

photon-upconversion nanoparticle; bioconjugation; immunoassay; prostate-specific antigen; protein p53; microtiter plate; massively parallel spectroscopy

Štítky

14313050, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 9. 5. 2025 10:11, doc. Mgr. Zdeněk Farka, Ph.D.

Anotace

V originále

Sensitive immunoassays for the detection of tumor biomarkers play an important role in the early diagnosis and therapy of cancer. Using luminescent nanomaterials as labels can significantly improve immunoassay performance, especially in terms of sensitivity. Lanthanide-doped photon-upconversion nanoparticles (UCNPs) are nanocrystals capable of converting near-infrared radiation into visible light, and their emission spectra can be tuned by altering the dopant ions. In this study, the bioconjugation between UCNPs and biomolecules was optimized, and different conjugates of Er- and Tm-doped UCNPs (NaYF4:Yb3+,Er3+ and NaYF4:Yb3+,Tm3+) were prepared for detecting prostate-specific antigen (PSA) and tumor protein p53, comparing heterogeneous and homogeneous assay formats. The heterogeneous sandwich immunoassay achieved detection limits of 1.3 pg mL−1 for PSA and 330 pg mL−1 for p53. The homogeneous immunoassays were based on massively parallel spectroscopy (MPS), a novel artificial intelligence-aided single-molecule approach, utilizing conjugates of two different monoclonal antibodies with Er- and Tm-doped UCNPs, respectively. The conjugates provide distinct emission spectra, with only sandwich immunocomplexes of analytes and both types of labels showing signals at the same location. MPS was suitable for observing the immunocomplexes in an aqueous dispersion using only a small sample volume. This innovative method achieved detection limits of 8.2 ng mL−1 and 390 pg mL−1 for PSA and p53, respectively. MPS eliminates the need for time-consuming washing steps required in heterogeneous immunoassays and is amenable to high-throughput applications.

Návaznosti

GA21-03156S, projekt VaV

Název: Foton-upkonverzní značky pro mikrofluidní jednomolekulové imunostanovení proteinových biomarkerů

Investor: Grantová agentura ČR, Foton-upkonverzní značky pro mikrofluidní jednomolekulové imunostanovení proteinových biomarkerů

Citovat

MAKHNEVA, Ekaterina; Pavel ŠPAČEK; Antonín HLAVÁČEK; Julie WEISOVÁ; Hans-Heiner GORRIS; Petr SKLÁDAL a Zdeněk FARKA. Bioconjugates of photon-upconversion nanoparticles with antibodies for the detection of prostate-specific antigen and p53 in heterogeneous and homogeneous immunoassays. Nanoscale. Royal Society of Chemistry, 2025, roč. 17, č. 18, s. 11401-11412. ISSN 2040-3364. Dostupné z: https://doi.org/10.1039/D5NR00176E.

@article{2495578,
   author = {Makhneva, Ekaterina and Špaček, Pavel and Hlaváček, Antonín and Weisová, Julie and Gorris, HansandHeiner and Skládal, Petr and Farka, Zdeněk},
   article_number = {18},
   doi = {https://doi.org/10.1039/D5NR00176E},
   keywords = {photon-upconversion nanoparticle; bioconjugation; immunoassay; prostate-specific antigen; protein p53; microtiter plate; massively parallel spectroscopy},
   language = {eng},
   issn = {2040-3364},
   journal = {Nanoscale},
   title = {Bioconjugates of photon-upconversion nanoparticles with antibodies for the detection of prostate-specific antigen and p53 in heterogeneous and homogeneous immunoassays},
   url = {https://pubs.rsc.org/en/content/articlelanding/2025/nr/D5NR00176E},
   volume = {17},
   year = {2025}
}

TY  - JOUR
ID  - 2495578
AU  - Makhneva, Ekaterina - Špaček, Pavel - Hlaváček, Antonín - Weisová, Julie - Gorris, Hans-Heiner - Skládal, Petr - Farka, Zdeněk
PY  - 2025
TI  - Bioconjugates of photon-upconversion nanoparticles with antibodies for the detection of prostate-specific antigen and p53 in heterogeneous and homogeneous immunoassays
JF  - Nanoscale
VL  - 17
IS  - 18
SP  - 11401-11412
EP  - 11401-11412
PB  - Royal Society of Chemistry
SN  - 20403364
KW  - photon-upconversion nanoparticle
KW  - bioconjugation
KW  - immunoassay
KW  - prostate-specific antigen
KW  - protein p53
KW  - microtiter plate
KW  - massively parallel spectroscopy
UR  - https://pubs.rsc.org/en/content/articlelanding/2025/nr/D5NR00176E
N2  - Sensitive immunoassays for the detection of tumor biomarkers play an important role in the early diagnosis and therapy of cancer. Using luminescent nanomaterials as labels can significantly improve immunoassay performance, especially in terms of sensitivity. Lanthanide-doped photon-upconversion nanoparticles (UCNPs) are nanocrystals capable of converting near-infrared radiation into visible light, and their emission spectra can be tuned by altering the dopant ions. In this study, the bioconjugation between UCNPs and biomolecules was optimized, and different conjugates of Er- and Tm-doped UCNPs (NaYF4:Yb3+,Er3+ and NaYF4:Yb3+,Tm3+) were prepared for detecting prostate-specific antigen (PSA) and tumor protein p53, comparing heterogeneous and homogeneous assay formats. The heterogeneous sandwich immunoassay achieved detection limits of 1.3 pg mL−1 for PSA and 330 pg mL−1 for p53. The homogeneous immunoassays were based on massively parallel spectroscopy (MPS), a novel artificial intelligence-aided single-molecule approach, utilizing conjugates of two different monoclonal antibodies with Er- and Tm-doped UCNPs, respectively. The conjugates provide distinct emission spectra, with only sandwich immunocomplexes of analytes and both types of labels showing signals at the same location. MPS was suitable for observing the immunocomplexes in an aqueous dispersion using only a small sample volume. This innovative method achieved detection limits of 8.2 ng mL−1 and 390 pg mL−1 for PSA and p53, respectively. MPS eliminates the need for time-consuming washing steps required in heterogeneous immunoassays and is amenable to high-throughput applications.
ER  -

MAKHNEVA, Ekaterina; Pavel ŠPAČEK; Antonín HLAVÁČEK; Julie WEISOVÁ; Hans-Heiner GORRIS; Petr SKLÁDAL a Zdeněk FARKA. Bioconjugates of photon-upconversion nanoparticles with antibodies for the detection of prostate-specific antigen and p53 in heterogeneous and homogeneous immunoassays. \textit{Nanoscale}. Royal Society of Chemistry, 2025, roč.~17, č.~18, s.~11401-11412. ISSN~2040-3364. Dostupné z: https://doi.org/10.1039/D5NR00176E.

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