Fluorescent excitation transfer immunoassay. A general method for determination of antigens.

摘要: A general immunochemical method for the assay of haptens and proteins has been devised applied to morphine, a morphine-albumin conjugate, human immunoglobulin G. fluorescein-labeled antigen quencher-labeled antibody are employed. By use fluorescein rhodamine as fluorescer quencher, respectively, dipole-dipole-coupled excitation energy transfer can occur within antigen-antibody complex. The resulting quenching fluorescence be inhibited by competitive binding with unlabeled antigen, Alternatively, separate samples labeled rhodamine, respectively. Unlabeled causes aggregation separately components resultant quenching. Using latter method, experiments suggest that up about 20 anti-morphine sites will associate conjugates. When an excess conjugate is present antibodies appear assemble in clumps on protein surface. Mathematical analysis morphine rhodamine-labeled gives approximate fit data, but calculations very dependent assumptions used.