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@article{2417985, author = {Sklenárová, Dorota and Hlaváček, Antonín and Křivánková, Jana and Brandmeier, Julian and Weisová, Julie and Řiháček, Michal and Gorris, HansandHeiner and Skládal, Petr and Farka, Zdeněk}, article_location = {Cambridge}, article_number = {June 2024}, doi = {http://dx.doi.org/10.1039/D4LC00346B}, keywords = {Photon-upconversion nanoparticle; Immunoassay; Microfluidics; Prostate-specific antigen}, language = {eng}, issn = {1473-0197}, journal = {Lab on a Chip}, title = {Single-molecule microfluidic assay for prostate-specific antigen based on magnetic beads and upconversion nanoparticles}, url = {https://pubs.rsc.org/en/content/articlelanding/2024/lc/d4lc00346b}, volume = {24}, year = {2024} }
TY - JOUR ID - 2417985 AU - Sklenárová, Dorota - Hlaváček, Antonín - Křivánková, Jana - Brandmeier, Julian - Weisová, Julie - Řiháček, Michal - Gorris, Hans-Heiner - Skládal, Petr - Farka, Zdeněk PY - 2024 TI - Single-molecule microfluidic assay for prostate-specific antigen based on magnetic beads and upconversion nanoparticles JF - Lab on a Chip VL - 24 IS - June 2024 SP - 3536-3545 EP - 3536-3545 PB - Royal Society of Chemistry SN - 14730197 KW - Photon-upconversion nanoparticle KW - Immunoassay KW - Microfluidics KW - Prostate-specific antigen UR - https://pubs.rsc.org/en/content/articlelanding/2024/lc/d4lc00346b N2 - Early-stage diagnosis of prostatic carcinoma is essential for successful treatment and, thus, significant prognosis improvement. In laboratory practice, the standard non-invasive diagnostic approach is the immunochemical detection of the associated biomarker, prostate-specific antigen (PSA). Ultrasensitive detection of PSA is essential for both diagnostic and recurrence monitoring purposes. To achieve exceptional sensitivity, we have developed a microfluidic device with a flow-through cell for single-molecule analysis using photon-upconversion nanoparticles (UCNPs) as a detection label. For this purpose, magnetic microparticles (MBs) were first optimized for the capture and preconcentration of PSA and then used to implement a bead-based upconversion-linked immunoassay (ULISA) in the microfluidic device. The digital readout based on counting single nanoparticle-labeled PSA molecules on MBs enabled a detection limit of 1.04 pg mL−1 (36 fM) in 50% fetal bovine serum, which is an 11-fold improvement over the respective analog MB-based ULISA. The microfluidic technique conferred several other advantages, such as easy implementation and the potential for achieving high-throughput analysis. Finally, it was proven that the microfluidic setup is suitable for clinical sample analysis, showing a good correlation with a reference electrochemiluminescence assay (recovery rates between 97% and 105%). ER -
SKLENÁROVÁ, Dorota, Antonín HLAVÁČEK, Jana KŘIVÁNKOVÁ, Julian BRANDMEIER, Julie WEISOVÁ, Michal ŘIHÁČEK, Hans-Heiner GORRIS, Petr SKLÁDAL a Zdeněk FARKA. Single-molecule microfluidic assay for prostate-specific antigen based on magnetic beads and upconversion nanoparticles. \textit{Lab on a Chip}. Cambridge: Royal Society of Chemistry, 2024, roč.~24, June 2024, s.~3536-3545. ISSN~1473-0197. Dostupné z: https://dx.doi.org/10.1039/D4LC00346B.
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