Patent ID: 6465259
Filing Date: 2002-10-15
Classification: C07D,G01N,Y10S,Y10T

Abstract:
A method for quantitatively and/or qualitatively assaying an analyte in a sample, with a non-radioactive assay, said analyte being a receptor binding compound, said method comprising the steps ofa) contacting the sample with material comprising a receptor for said analyte in order for receptor-analyte binding to occur and b) further contacting the sample with a non-radioactive detectable ligand for the receptor in order for receptor-ligand binding to occur, followed by c) separating the resulting receptor bound and free fractions, d) subjecting the receptor bound fraction to dissociating conditions releasing the non-radioactive ligand from the receptor, e) assaying for the dissociated non-radioactive ligand so as to detect the detectable ligand, and f) determining presence or amount of said analyte from results of step (e); said receptor being present in a concentration between 0.5-5 nM, said non-radioactive detectable ligand being at least 90% pure, said non-radioactive detectable ligand being non radioactively detectable, said detectable ligand being added in an amount corresponding to the amount required for occupation by the ligand of 10-75% of the receptor material present in the assay in absence of analyte at a receptor concentration below the Kd of the ligand and receptor under conditions otherwise corresponding to those of the assay, wherein Kd is the equilibrium dissociation constant of the detectable ligand, said dissociating conditions and said non-radioactive detectable ligand being such that at least 90% bound ligand is released, in such a form that less than 10% of the resulting detectable product reassociates with the receptor, said dissociating conditions and said non-radioactive detectable ligand being selected such that the resulting dissociated ligand is quantitatively and/or qualitatively detectable, wherein when said method effects a quantitative assay of said analyte, said method further comprises determining analyte content in relation to calibration curves.