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Phycobiliproteins as Labels in Immunoassay

  • Mel N. Kronick

Abstract

Fluorescence has for many years been viewed as a potential means of providing high sensitivity in a non-isotopic test format (Soini and Hemmila, 1979; Hemmila, 1985). As Jolley et al. (1984) have clearly pointed out, the number of photons emitted from a sample of fluorescently tagged molecules can exceed the number of gamma ray photons emitted from the same number of gamma-ray-emitting tagged molecules. The reason for this is two fold: each radioactive molecule can emit only once and it emits its photon only when it naturally and spontaneously decays. Nevertheless, high sensitivity assays using fluorescence have not replaced radioactive assays to any great extent. Non-isotopic versions of most high sensitivity assays are run today using enzymes as labels. Fluorescence immunoassay has penetrated the marketplace but only in certain applications. One technique, fluorescence polarization (Dandliker and Saussure, 1970; Jolley et al., 1981), has proven very useful, but only for small molecules such as drugs which are present at concentrations from approximately 10−6 to 10−10 M. The dynamic range inherent in fluorescence polarization assays in a competition assay configuration result in this limitation and the physics of fluorescence polarization prevents its being adapted easily to immunometric assays or even to competition assays for larger (≳1000 molecular weight) molecules. Assays based upon fluorescence quenching or fluorescence energy transfer (Ullman et al., 1976; Ullman and Khanna, 1981) similarly have proven most useful to date only in competition assays for analytes of concentration greater than about 10−10 M.

Keywords

Fluorescence Polarization Complete Amino Acid Sequence Fluorescence Immunoassay High Sensitivity Assay Porphyridium Cruentum 
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Copyright information

© Plenum Press, New York 1988

Authors and Affiliations

  • Mel N. Kronick
    • 1
  1. 1.Applied BiosystemsFoster CityUSA

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