Preparation of 125I-labeled human growth hormone of high quality binding properties endowed with long-term stability.
125I-labeled human growth hormone (125I-labeled.hGH) was prepared by using two variants of the chloramine T labelling procedure and purified by polyacrylamide gel electrophoresis (PAGE) of the reaction mixture. Variant A produced a tracer with high specific activity (100 +/- 10 microCi/microgram), high maximal binding capacity to antibodies (93%) and long-term stability (at least 150 days at -20 degrees C). No diiodinated tyrosil residues could be detected in this tracer. Variant B was devised to obtain higher yields of labeled hormone. The electrophoresis of the iodination mixture revealed two radioactive components with Rm values of 0.49 and 0.55 which result from the iodination of hGH variants preexisting in the starting material. Both tracers had similar specific activities (70 +/- 10 microCi/microgram), high maximal binding capacity to antibodies or receptors (80-100%, after 80 days of their obtention) and high stability (at least 100 days at -20 degrees C). It is concluded that the iododerivatives of hGH obtained by either method are adequate to perform radioimmunoassay and receptor studies and have long-term stability.
Biscayart
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PL
Instituto de Quimica y Fisicoquímica Biológicas (UBA-CONICET) Facultad de Farmacia y Bioquimica, Buenos Aires, Argentina.
Paladini
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AC
Vita
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Roguin
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LP
D002700:Chloramines; D004105:Diiodotyrosine / Q000032:analysis; D004355:Drug Stability; D004591:Electrophoresis, Polyacrylamide Gel; D013006:Growth Hormone / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D007457:Iodine Radioisotopes; D007470:Monoiodotyrosine / Q000032:analysis; D011863:Radioimmunoassay; D011869:Radioligand Assay; D014105:Tosyl Compounds
D002700:Chloramines; D007457:Iodine Radioisotopes; D014105:Tosyl Compounds; C016300:chloramine-T; D004105:Diiodotyrosine; D013006:Growth Hormone; D007470:Monoiodotyrosine