Influence of erythrocyte iodothyronine-binding proteins on radioimmunoassay of thyroxin in dried blood spots.
Three erythrocyte proteins, one identified as hemoglobin, bind thyroid hormones. Using a dextran/charcoal radioimmunoassay for thyroxin in dried blood spots, we demonstrate that such binding differs with the buffer used. Barbital, phosphate, and borate buffers significantly enhance the binding more than glycine and tris(hydroxymethyl)methylamine buffers. Binding is not affected by agents commonly used to inhibit thyroxin binding to serum proteins. A highly significant nonlinear direct relationship between sample storage (temperature and duration) and increased thyroxin-erythrocyte binding is documented, together with an associated decrease in assayed concentrations of thyroxin. However, concomitant serial measurement of thyroxin with polyethylene glycol and combined double-antibody/polyethylene glycol radioimmunoassays produced no evidence of interference by erythrocyte proteins in the radioimmune reaction. We conclude that erythrocyte proteins act only as low-affinity secondary binders in radioimmunoassay for thyroxin.
D001793:Blood Preservation / Q000379:methods*; D002021:Buffers; D002352:Carrier Proteins / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism*; D006801:Humans; D011863:Radioimmunoassay; D012016:Reference Values; D013696:Temperature; D013974:Thyroxine / Q000097:blood*; D013997:Time Factors; D014284:Triiodothyronine / Q000378:metabolism*
D002021:Buffers; D002352:Carrier Proteins; D014284:Triiodothyronine; D013974:Thyroxine