Microencapsulated antibodies in radioimmunoassay. III. Determination of cortisol.

We describe a method for directly measuring total plasma cortisol by use of a microencapsulated antibody. The antibody microcapsules were incubated with plasma and 125I-labeled cortisol in a competitive reaction at 37 degrees C for 15 min, then separated by centrifugation; the radioactivity of the pellet was counted for 1 min. Analytical recovery of cortisol from three plasma pools was 97.6% (SD 8.5%) and was unaffected by triglyceridemia, hemolysis, or icterus. The standard curve was linear to 500 microgram/L and the sensitivity was 10 microgram/l. Recovery of cortisol added as the hemisuccinate was 102% (SD 10.6%), whereas that of the conjugate cortisol hemisuccinate/bovine serum albumin was 4.3% (SD 3.5%). This confirmed that the microcapsule excludes large molecules. The within-day CV for two control pools was 9.8 and 12.8%, the day-to-day variation 13.4 and 13.8%.

Microencapsulated antibodies in radioimmunoassay. II. Determination of free thyroxine.

We describe a method for directly measuring free thyroxine in human serum. Antibody to thyroxine was encapsulated within a semipermeable nylon membrane that excludes substances of relative molecular mass greater than 20,000. The microencapsulated antibody was then presaturated with [125I]thyroxine. Serum incubated with the microcapsules initiated a displacement reaction between free thyroxine and [125I]thyroxine bound to the antibody. The displaced thyroxine was separated from the bound thyroxine by centrifugation and the concentration of free thyroxine determined from a standard curve prepared by use of known amounts of free thyroxine. The within-day coefficient of variation for two control samples was 8.8 and 6.8%; day-to-day precision for the same material was 12.5 and 13.5%. Lipemia, icterus, or hemoglobin had no adverse effect. Interlaboratory evaluation of the procedure revealed no significant difference in mean values for 20 speciments (p greater than 0.05).

Microencapsulated antibodies in radioimmunoassay--I. Determination of digoxin.

We describe the application of the microencapsulated-antibody technique to the radioimmunoassay of digoxin in serum. Droplets of emulsified rabbit antibody are microencapsulated in a semipermeable nylon membrane by an interfacial polymerization technique. The antibody microcapsules are incubated with 125I-labeled digoxin and unlabeled digoxin for 15 min at 37 degrees C, then free and bound digoxin are separated by centrifugation. Subtherapeutic, therapeutic, and toxic concentrations of digoxin in sera can be determined, with use of a standard curve prepared by use of known amounts of digoxin. With this technique we obtained an intra-laboratory correlation coefficient of 0.945 for 100 patients' sera and one of 0.940 for interlaboratory results for 21 sera (10 laboratories) when compared to a routine clinical laboratory radioimmunoassay for digoxin. Icterus, lipemia, hemoglobin, or disproteinemia had no effect on the analytical recovery of digoxin. The standard curve was linear to 6 microgram/L; the sensitivity was 0.25 microgram/L.