Influence of Label and Solid Support on the Performance of Heterogeneous Immunoassays

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Authors

MAKHNEVA Ekaterina SKLENÁROVÁ Dorota BRANDMEIER Julian HLAVÁČEK Antonín GORRIS Hans-Heiner SKLÁDAL Petr FARKA Zdeněk

Year of publication 2022
Type Article in Periodical
Magazine / Source Analytical Chemistry
MU Faculty or unit

Faculty of Science

Citation
Web https://pubs.acs.org/doi/10.1021/acs.analchem.2c03543
Doi http://dx.doi.org/10.1021/acs.analchem.2c03543
Keywords Immunoassay; Magnetic preconcentration; Photon-upconversion nanoparticle; Prostate-specific antigen
Description Conventional immunochemical methods used in clinical analysis are often not sensitive enough for early-stage diagnosis, resulting in the need for novel assay formats. Here, we provide a detailed comparison of the effect of different labels and solid supports on the performance of heterogeneous immunoassays. When comparing three types of streptavidin-modified labels¦horseradish peroxidase, carboxyfluorescein, and photon-upconversion nanoparticles (UCNPs)¦UCNPs led to the most sensitive and robust detection of the cancer biomarker prostate-specific antigen. Additionally, we compared the immunoassay formats based on conventional microtiter plates and magnetic microbeads (MBs). In both cases, the highest signal-to-background ratios and the lowest limits of detection (LODs) were obtained by using the UCNP labels. The MB-based upconversion-linked immunosorbent assay carried out with a preconcentration step provided the lowest LOD of 0.46 pg/mL in serum. The results demonstrate that the use of UCNPs and MBs can significantly improve the sensitivity and working range of heterogeneous immunoassays for biomarker detection.
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