Homogeneous fluorassay methods employing fluorescent background rejection and water-soluble rare earth metal chelates

Abstract

Homogeneous assays for determining quantitatively the extent of a specific binding reaction can be carried out effectively on very dilute solutions using measurements of fluorescence if a fluorescence measurement scheme that is capable of rejecting short-lived background fluorescence is employed and if the fluorescent group being measured has the following properties: a. the group being measured must be a rare earth metal chelate complex combination; b. the chelate must be water-soluble; c. the complex combination must also be stable in extremely dilute aqueous solutions, that is, the measured chelate must have at least one ligand having a metal-to-ligand binding constant of at least about 10,M,or greater and it must have a fluorescent emission that is long-lived compared to the longest decay lifetime of ambient substances and have a half life of from 0.01 to 50 msec.