OPTIMIZATION OF RADIOLIGAND BINDING AND RADIOIMMUNO ASSAYS

Radioligand binding assay (RLA) and radioimmunoassay (RIA) development involves consideration of a large number variables which influence si gnal readout or system errors, with contrary effects on assay performa nce. We have developed an optimization strategy according to statistic al criteria which incorporates these variables and which specifies a m inimum data set for a systematic analysis. Increased background and er ror fractions are seen to reduce assay power, whereas increased specif ic activity, affinity, and counting time increase power. The optimal c oncentration of radiolabeled developer in the test is dependent upon a balance of these factors, but in general yields a broad optimum, Thes e analyses suggest several strategies available to the investigator fo r improving assay performance. The optimization method is adapted to s tandard microcomputer spreadsheet formats which allow ready applicatio n of these procedures in any research laboratory. (C) 1906 Academic Pr ess, Inc.