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Multi-wavelength immunoassays using surface plasmon-coupled emission.

Evgenia Matveeva1, Joanna Malicka, Ignacy Gryczynski

  • 1Department of Biochemistry and Molecular Biology, Center for Fluorescence Spectroscopy, University of Maryland at Baltimore, 725 West Lombard Street, Baltimore, MD 21201, USA.

Biochemical and Biophysical Research Communications
|December 31, 2003
PubMed
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We developed a novel multi-wavelength immunoassay technique using surface plasmon-coupled emission (SPCE). This method enables independent detection of multiple analytes by utilizing distinct emission angles for each wavelength, enhancing diagnostic capabilities.

Area of Science:

  • Biotechnology
  • Analytical Chemistry
  • Optics

Background:

  • Immunoassays are crucial for detecting biomolecules.
  • Current methods can be limited in multiplexing capabilities.
  • Surface plasmon-coupled emission (SPCE) offers unique optical properties.

Purpose of the Study:

  • To introduce a new multi-wavelength immunoassay method.
  • To leverage SPCE for simultaneous detection of multiple analytes.
  • To demonstrate the feasibility of SPCE for advanced diagnostics.

Main Methods:

  • Utilized surface plasmon-coupled emission (SPCE) with a silver film.
  • Coupled excited fluorophores (Rhodamine Red-X, AlexaFluor 647) to the metal film.
  • Directed emission into a glass substrate at wavelength-dependent angles.

Related Experiment Videos

  • Immobilized target antigen protein on the silver surface.
  • Used labeled antibodies for antigen detection.

    • Main Results:

    • Demonstrated independent angular emission for different wavelengths.
    • Successfully detected two different antibodies against a surface-bound antigen.
    • Achieved distinct angular separation of emission signals.
    • Showcased the potential for multiplexing 4+ analytes.

    Conclusions:

    • SPCE immunoassays provide a robust platform for multi-analyte detection.
    • The angular emission property allows for precise, independent measurements.
    • This technique offers a scalable approach for developing advanced diagnostic tools.