Method of epitope scanning using fluorescence polarization
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United States of America Patent
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Issued Date -
Sep 23, 2004
app pub date -
Mar 20, 2003
filing date -
Mar 20, 2003
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Abstract
An antigenic protein includes a known amino acid sequence. To locate one or more epitopes of the antigenic protein, a plurality of distinct peptides are synthesized bound to respective solid-phase supports via selectively cleavable linkers. Each of the distinct peptides corresponds to a sub-sequence of the antigenic protein's known amino acid sequence. While the peptides are bound to their respective supports, they are conjugated to a fluorophore. The conjugated peptides are then selectively cleaved from their supports, and the fluorescence polarization of the free conjugated peptides is measured. The free conjugated peptides are each combined with an antibody that is able to bind to the antigenic protein, and the fluorescence polarization of the mixtures is measured. A substantial increase in fluorescence polarization of a mixture indicates the presence of an epitope.
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