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Sensitive bioaffinity assays with individual microparticles and time-resolved fluorometry

T Lövgren1, P Heinonen, P Lehtinen

  • 1Department of Biotechnology, University of Turku, Finland. timo.lovgren@utu.fi

Clinical Chemistry
|October 28, 1997
PubMed
Summary
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Future immunoassays and nucleic acid assays will use miniaturized formats. Fluorescent europium chelates and time-resolved microfluorometry offer sensitive detection for individual microparticles, advancing diagnostic technologies.

Area of Science:

  • Biotechnology
  • Analytical Chemistry
  • Molecular Diagnostics

Background:

  • Miniaturized assays on microchips or microparticles require sensitive, spatially resolved detection technologies.
  • Existing detection methods may lack the sensitivity needed for single-particle analysis in miniaturized formats.

Purpose of the Study:

  • To evaluate fluorescent lanthanide chelates and time-resolved microfluorometry as a next-generation detection technology for miniaturized immunoassays and nucleic acid hybridization assays.
  • To demonstrate the capability of this technology for detecting specific analytes at low concentrations on individual microparticles.

Main Methods:

  • Utilized fluorescent europium chelates as labels for detection.
  • Employed time-resolved microfluorometry for sensitive signal detection with spatial resolution.

Related Experiment Videos

  • Developed miniaturized noncompetitive immunoassays for prostate-specific antigen (PSA) and hybridization assays for cystic fibrosis deltaF508 mutation.

    • Main Results:

    • Successfully detected 11,000 europium molecules on individual microparticles.
    • Quantitatively detected 5 ng/L (0.05 amol per particle) of PSA with excellent precision (CV <10%) in a miniaturized immunoassay.
    • Detected the deltaF508 mutation for cystic fibrosis on individual microparticles using a hybridization assay.

    Conclusions:

    • Fluorescent lanthanide chelate labels combined with time-resolved microfluorometry provide a highly sensitive and precise detection method for miniaturized assays.
    • This technology is suitable for detecting biomarkers like PSA and genetic mutations like deltaF508 at the single-particle level.
    • This approach represents a significant advancement for future immunoassays and nucleic acid hybridization assays in miniaturized formats.