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).