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

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 immun...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2022-11, Vol.94 (47), p.16376-16383
Main Authors: Makhneva, Ekaterina, Sklenárová, Dorota, Brandmeier, Julian C., Hlaváček, Antonín, Gorris, Hans H., Skládal, Petr, Farka, Zdeněk
Format: Article
Language:English
Subjects:
Antigens
Biomarkers
Chemistry
Horseradish peroxidase
Humans
Immunoassay
Immunoassay - methods
Immunoassays
Immunosorbents
Labels
Limit of Detection
Magnetics
Male
Microspheres
Nanoparticles
Peroxidase
Prostate cancer
Prostate-specific antigen
Streptavidin
Upconversion
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Summary: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.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.2c03543