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Proceedings Paper

Pi overlapping ring systems contained in a homogeneous assay: a novel homogeneous assay for antigens
Author(s): David A. Kidwell
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Paper 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.

Paper Details

Date Published: 18 May 1993
PDF: 12 pages
Proc. SPIE 1885, Advances in Fluorescence Sensing Technology, (18 May 1993); doi: 10.1117/12.144741
Show Author Affiliations
David A. Kidwell, Naval Research Lab. (United States)

Published in SPIE Proceedings Vol. 1885:
Advances in Fluorescence Sensing Technology
Joseph R. Lakowicz; Richard B. Thompson, Editor(s)

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