Heterogeneity of 125I-labeled human thyroglobulin preparations.
This study was undertaken to evaluate the heterogeneity and stability of 125I-labeled human thyroglobulin (Tg) tracers. Tg, labeled with 125I by either a Chloramine T (CT) or a Glucose Oxidase/Lactoperoxidase (GO) method, showed considerable heterogeneity of labeled components, the relative proportions of which were a function of the Tg preparation and the iodination method used. The four largest components had apparent molecular weights as follows: A = 1 200 000 Da; B = 670 000 Da; C = 530 000 Da and D = 290 000 Da. Both B and C were immunoactive. B was considered to be 19S Tg. A non-specific binding difference, (NSB delta) between nonhuman matrices used for diluting standards and human sera was found for the partly immunoactive aggregate component A, (5-20%) and the nonimmunoactive component D, (20-50%), but was minimally present for components B and C (less than 5%). The [125I]19S Tg(B), stored at -70 degrees C, showed rapid spontaneous decomposition with time (50% lost by 8 days), with generation of C, D and iodide. The loss of B was related to specific activity and was least in GO labels. 125I labeling of Tg by GO produced tracers with better immunoactivity, stability and lower NSB delta than comparative CT tracers. Definitive purification and repurification of [125I]Tg tracers before use is necessary in order to remove degradation products with the potential to compromise the accuracy and specificity of serum Tg radioimmunoassay (RIA) measurement.
D002845:Chromatography / Q000379:methods; D002850:Chromatography, Gel; D003208:Concanavalin A; D004586:Electrophoresis / Q000379:methods; D006801:Humans; D007120:Immunochemistry; D007457:Iodine Radioisotopes; D007553:Isotope Labeling; D008970:Molecular Weight; D013954:Thyroglobulin / Q000032:analysis*
D007457:Iodine Radioisotopes; D003208:Concanavalin A; D013954:Thyroglobulin