Pi overlapping ring systems contained in a homogeneous assay: a novel homogeneous assay for antigens

David A. Kidwell

doi:10.1117/12.144741

18 May 1993 Pi overlapping ring systems contained in a homogeneous assay: a novel homogeneous assay for antigens

David A. Kidwell

Proceedings Volume 1885, Advances in Fluorescence Sensing Technology; (1993) https://doi.org/10.1117/12.144741
Event: OE/LASE'93: Optics, Electro-Optics, and Laser Applications in Scienceand Engineering, 1993, Los Angeles, CA, United States

Abstract

A novel immunoassay, Pi overlapping ring systems contained in a homogeneous assay (PORSCHA), is described. This assay relies upon the change in fluorescent spectral properties that pyrene and its derivatives show with varying concentration. Because antibodies and other biomolecules can bind two molecules simultaneously, they can change the local concentration of the molecules that they bind. This concentration change may be detected spectrally as a change in the fluorescence emission wavelength of an appropriately labeled biomolecule. Several tests of PORSCHA have been performed which demonstrate this principle. For example: with streptavidin as the binding biomolecule and a biotin labeled pyrene derivative, the production of the excimer emitting at 470 nm is observed. Without the streptavidin present, only the monomer emitting at 378 and 390 nm is observed. The ratio of monomer to excimer provides the concentration of unlabeled biotin in the sample. Approximately 1 ng/mL of biotin may be detected with this system using a 50 (mu) l sample (2 X 10^-16 moles biotin). The principles behind PORSCHA, the results with the streptavidin/biotin system are discussed and extensions of the PORSCHA concept to antibodies as the binding partner and DNA in homogeneous assays are suggested.

Citation Download Citation

David A. Kidwell "Pi overlapping ring systems contained in a homogeneous assay: a novel homogeneous assay for antigens", Proc. SPIE 1885, Advances in Fluorescence Sensing Technology, (18 May 1993); https://doi.org/10.1117/12.144741

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