Background-free detection of prostate-specific antigen
utilizing an immunomagnetic assay with upconversion
nanoparticles

a 2024

Background-free detection of prostate-specific antigen utilizing an immunomagnetic assay with upconversion nanoparticles

SKLENÁROVÁ, Dorota; Ekaterina MAKHNEVA; Julian BRANDMEIER; Antonín HLAVÁČEK; Hans-Heiner GORRIS et al.

Základní údaje

Originální název

Background-free detection of prostate-specific antigen utilizing an immunomagnetic assay with upconversion nanoparticles

Autoři

SKLENÁROVÁ, Dorota; Ekaterina MAKHNEVA; Julian BRANDMEIER; Antonín HLAVÁČEK; Hans-Heiner GORRIS a Zdeněk FARKA

Vydání

UPCON 2024, 2024

Další údaje

Jazyk

angličtina

Typ výsledku

Konferenční abstrakt

Obor

10406 Analytical chemistry

Stát vydavatele

Kanada

Utajení

není předmětem státního či obchodního tajemství

Označené pro přenos do RIV

Organizační jednotka

Přírodovědecká fakulta

Příznaky

Mezinárodní význam

Změněno: 10. 2. 2025 11:39, doc. Mgr. Zdeněk Farka, Ph.D.

Anotace

V originále

Prostatic carcinoma is a severe oncologic disease that can be diagnosed via the immunochemical detection of its most important biomarker, the prostate-specific antigen (PSA). Blood serum concentration of PSA up to 2.5 ng/mL is considered normal and can be generally detected by conventional techniques, such as the enzyme-linked immunosorbent assay (ELISA). However, more sensitive approaches are necessary for early-stage diagnosis and minor changes monitoring. Photon-upconversion nanoparticles (UCNPs) are lanthanide-doped nanocrystals that exhibit anti-Stokes luminescence, thus omitting optical background interference and enhancing detection sensitivity. It is possible to utilize them as sensitive immunoassay labels in, e.g., the microtiter plate upconversion-linked immunosorbent assay (ULISA). However, even the sensitivity of the ULISA may not sufficient for the detection of extremely low-abundant analytes. Magnetic microparticles (MBs) are a promising alternative solid phase for microtiter plate immunoassays, allowing for analyte preconcentration and improving the assay sensitivity. We have developed and optimized a sandwich immunoassay for the detection of PSA based on MBs and UCNPs. We conjugated a monoclonal anti-PSA antibody to MBs and a polyclonal one to UCNPs via streptavidin-biotin binding and tested their functionality. Optimal concentrations of MBs and serum were selected, followed by the fully optimized analysis of model samples of spiked serum. The assay in serum reached an LOD of 11.1 pg/mL, comparable to an ELISA using identical immunoreagents. An additional magnetic preconcentration step further improved the LOD to 0.25 pg/mL, which was 3-fold better than ELISA and 44-fold better than MB-based ELISA using identical immunoreagents. The developed technique was then performed in clinical sample analysis, demonstrating the potential of using MBs combined with UCNPs for the sensitive detection of cancer biomarkers. Moreover, we are currently aiming to optimize the method to reach ultrasensitive single-molecule detection in a microfluidic setup as a means to enhance the sensitivity further.

Návaznosti

MUNI/A/1582/2023, interní kód MU

Název: Podpora biochemického výzkumu v roce 2024

Investor: Masarykova univerzita, Podpora biochemického výzkumu v roce 2024

Citovat

SKLENÁROVÁ, Dorota; Ekaterina MAKHNEVA; Julian BRANDMEIER; Antonín HLAVÁČEK; Hans-Heiner GORRIS a Zdeněk FARKA. Background-free detection of prostate-specific antigen utilizing an immunomagnetic assay with upconversion nanoparticles. In UPCON 2024. 2024.

@proceedings{2475457,
   author = {Sklenárová, Dorota and Makhneva, Ekaterina and Brandmeier, Julian and Hlaváček, Antonín and Gorris, HansandHeiner and Farka, Zdeněk},
   booktitle = {UPCON 2024},
   language = {eng},
   title = {Background-free detection of prostate-specific antigen utilizing an immunomagnetic assay with upconversion nanoparticles},
   year = {2024}
}

TY  - CONF
ID  - 2475457
AU  - Sklenárová, Dorota - Makhneva, Ekaterina - Brandmeier, Julian - Hlaváček, Antonín - Gorris, Hans-Heiner - Farka, Zdeněk
PY  - 2024
TI  - Background-free detection of prostate-specific antigen utilizing an immunomagnetic assay with upconversion nanoparticles
N2  - Prostatic carcinoma is a severe oncologic disease that can be diagnosed via the immunochemical detection of its most important biomarker, the prostate-specific antigen (PSA). Blood serum concentration of PSA up to 2.5 ng/mL is considered normal and can be generally detected by conventional techniques, such as the enzyme-linked immunosorbent assay (ELISA). However, more sensitive approaches are necessary for early-stage diagnosis and minor changes monitoring. Photon-upconversion nanoparticles (UCNPs) are lanthanide-doped nanocrystals that exhibit anti-Stokes luminescence, thus omitting optical background interference and enhancing detection sensitivity. It is possible to utilize them as sensitive immunoassay labels in, e.g., the microtiter plate upconversion-linked immunosorbent assay (ULISA). However, even the sensitivity of the ULISA may not sufficient for the detection of extremely low-abundant analytes. Magnetic microparticles (MBs) are a promising alternative solid phase for microtiter plate immunoassays, allowing for analyte preconcentration and improving the assay sensitivity. We have developed and optimized a sandwich immunoassay for the detection of PSA based on MBs and UCNPs. We conjugated a monoclonal anti-PSA antibody to MBs and a polyclonal one to UCNPs via streptavidin-biotin binding and tested their functionality. Optimal concentrations of MBs and serum were selected, followed by the fully optimized analysis of model samples of spiked serum. The assay in serum reached an LOD of 11.1 pg/mL, comparable to an ELISA using identical immunoreagents. An additional magnetic preconcentration step further improved the LOD to 0.25 pg/mL, which was 3-fold better than ELISA and 44-fold better than MB-based ELISA using identical immunoreagents. The developed technique was then performed in clinical sample analysis, demonstrating the potential of using MBs combined with UCNPs for the sensitive detection of cancer biomarkers. Moreover, we are currently aiming to optimize the method to reach ultrasensitive single-molecule detection in a microfluidic setup as a means to enhance the sensitivity further.
ER  -

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