Mini-RIA: adaptation of conventional 125I-labeled radioimmunoassay to a 96-tube format.
Radioimmunoassay (RIA) employing iodinated ligands represents a popular measurement method for small molecules due to its excellent sensitivity and specificity. Yet performing RIAs of large numbers of tubes remains a tedious laboratory chore due to the need to individually handle tubes multiple times. We here present a method in which conventional 125I-labeled RIA ([125I] RIA) is adapted to a microtiter plate format, termed mini-RIA. Tubes are handled in batch for centrifugation or during the separation of antibody-bound ligand from free ligand. A simple draining device for batch decantation of free ligand from 96 minitubes is used. Optimal conditions for the mini-RIA were established using two workup methods-double-antibody immunoprecipitation and direct polyethylene glycol precipitation. Use of the mini-RIA method was found to result in a considerable savings in assay times; in addition, the sensitivity of the mini-RIA was improved over conventional RIA. The mini-RIA is particularly useful for assay of large numbers of samples derived from chromatographic methods, since aliquots can be transferred directly from the fraction collector into the minitubes using multiple channel pipettors. Because the method is flexible with regard to assay workup, we predict that most conventional [125I] RIAs can be adapted to the mini-RIA format.
Laurent
Virginie
V
Department of Biochemistry and Molecular Biology, Louisiana State University Health Sciences Center, 1901 Perdido Street, New Orleans, LA 70112, USA.
Lindberg
Iris
I
D000324:Adrenocorticotropic Hormone / Q000276:immunology; D000906:Antibodies / Q000032:analysis; D011232:Chemical Precipitation; D002850:Chromatography, Gel / Q000379:methods; D007457:Iodine Radioisotopes / Q000737:chemistry*; D008904:Miniaturization / Q000295:instrumentation; D011092:Polyethylene Glycols / Q000737:chemistry; D011863:Radioimmunoassay / Q000295:instrumentation / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D005719:gamma-Globulins / Q000737:chemistry / Q000276:immunology
D000906:Antibodies; D007457:Iodine Radioisotopes; D005719:gamma-Globulins; D011092:Polyethylene Glycols; D000324:Adrenocorticotropic Hormone