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Upconversion Nanoparticle-Based Dot-Blot Immunoassay for Quantitative Biomarker Detection.

Jakub Máčala1, Ekaterina Makhneva1, Antonín Hlaváček2

  • 1Department of Biochemistry, Faculty of Science, Masaryk University, Kamenice 5, 625 00 Brno, Czech Republic.

Analytical Chemistry
|June 13, 2024
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Summary
This summary is machine-generated.

Photon-upconversion nanoparticles (UCNPs) significantly enhance dot-blot immunoassay sensitivity for detecting low-abundance biomarkers. This UCNP-based assay offers improved quantitative analysis for clinical diagnostics.

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Area of Science:

  • Biomedical Engineering
  • Nanotechnology
  • Analytical Chemistry

Background:

  • Dot-blot immunoassays are common for biomarker detection but often lack sensitivity for quantitative analysis.
  • Existing assays are typically limited to qualitative or semi-quantitative measurements.
  • Optical background interference can hinder sensitive detection in traditional assays.

Purpose of the Study:

  • To develop a highly sensitive dot-blot immunoassay using photon-upconversion nanoparticles (UCNPs) for quantitative biomarker detection.
  • To overcome the sensitivity limitations of conventional dot-blot assays.
  • To demonstrate the versatility and robustness of UCNP-based detection for various clinical biomarkers.

Main Methods:

  • Optimization of a dot-blot immunoassay on nitrocellulose membranes using UCNPs as labels.
  • Quantitative analysis of human serum albumin (HSA) to establish assay performance metrics.
  • Comparison of UCNP labels against commercial quantum dots for sensitivity and signal-to-background ratio.
  • Validation of the assay using spiked urine and serum samples for prostate-specific antigen (PSA) and cardiac troponin (cTn).
  • Analysis of clinical patient samples and correlation with electrochemiluminescence immunoassay (ECLIA) results.

Main Results:

  • The UCNP-based dot-blot assay achieved a limit of detection (LOD) of 0.19 ng/mL for HSA with a signal-to-background ratio (S/B) of 722.
  • UCNPs demonstrated superior performance compared to quantum dots, with an LOD of 4.32 ng/mL and S/B of 3.
  • The assay successfully detected PSA (LOD 9.4 pg/mL) and cTn (LOD 0.62 ng/mL) in spiked serum samples.
  • Analysis of clinical samples showed strong correlation with ECLIA, with recovery rates ranging from 89% to 117%.

Conclusions:

  • Photon-upconversion nanoparticles serve as highly sensitive labels for dot-blot immunoassays.
  • UCNP-based dot-blot assays enable quantitative detection of low-abundance biomarkers.
  • This technology offers a promising platform for sensitive and robust clinical diagnostics.