Patent ID: 6207397
Filing Date: 2001-03-27
Classification: G01N

Abstract:
An in vitro assay method for identifying a test substance which inhibits the mutual association of two molecules, said method comprising:(a) providing a first molecule and a second molecule capable of mutual association, said second molecule bearing a covalently linked fluorophore,(b) preparing a mixture containing said first and second molecules and at least one test substance,(c) irradiating said mixture with polarized light of a suitable wavelength permitting excitation of the fluorophore as indicated by emission of polarized light,(d) measuring the degree of polarization of the emission, and(e) determining the effect of the presence or concentration of the test substance on the observed emission polarization of the mixture, wherein inhibitory activity of said test substance correlates with decreased depolarization values, and wherein the first molecule comprises a polypeptide which contains at least one receptor domain selected from the group consisting of a Src Homology region (""SH"") 2 domain SH3 domain, Phosphotyrosine Interaction domain (""PID"") or WW domain.