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Mass spectrometric immunoassay.

R W Nelson1, J R Krone, A L Bieber

  • 1Department of Chemistry and Biochemistry, Arizona State University, Tempe, 85287-1604 USA.

Analytical Chemistry
|April 1, 1995
PubMed
Summary
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This study introduces a novel immunoassay combining immunoaffinity capture with mass spectrometry for sensitive and specific antigen detection. This method enables rapid screening of complex biological samples for multiple toxins.

Area of Science:

  • Analytical Chemistry
  • Biochemistry
  • Immunology

Background:

  • Complex biological mixtures often cause signal suppression in traditional matrix-assisted laser desorption/ionization mass spectrometry.
  • Accurate detection and quantification of specific antigens in biological samples present significant analytical challenges.

Purpose of the Study:

  • To develop a general immunoassay method for sensitive and specific antigen identification and quantification.
  • To overcome limitations of traditional mass spectrometry techniques for complex biological samples.
  • To enable simultaneous screening for multiple antigens within a single assay.

Main Methods:

  • Microscale immunoaffinity capture of target antigens.
  • Mass-specific identification and quantification using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS).

Related Experiment Videos

  • Analyte concentration and reduction of signal suppression through immunoaffinity capture.
  • Quantification using a single antibody to capture both antigen and a mass-modified variant.

    • Main Results:

    • The developed immunoassay effectively overcomes signal suppression in complex mixtures.
    • Mass spectrometric detection provides unambiguous, artifact-resistant antigen identification.
    • The method allows for the screening of multiple, mass-resolved antigens in a single assay.
    • Subnanomolar sensitivities were achieved with analysis times under 1 hour.
    • Successful application in screening human blood for specific snake venom toxins (myotoxin a and Mojave toxin).

    Conclusions:

    • This novel immunoassay offers a rapid, selective, and quantitative approach for antigen detection.
    • The combination of immunoaffinity capture and MALDI-TOF MS provides enhanced sensitivity and specificity.
    • The method is versatile for screening complex biological systems for multiple analytes.