Evaluation of a radioimmunoassay for testosterone estimation

• Published in: Journal of steroid biochemistry Vol. 4; no. 6; pp. 665 - 676
• Main Authors: Verjans, H.L., Cooke, B.A., De Jong, F.H., De Jong, C.M.M., Van Der Molen, H.J.
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 01.11.1973
• Subjects: Animals; Cattle; Charcoal; Chromatography; Chromatography, Gas; Computers; Dextrans; Evaluation Studies as Topic; Humans; Male; Methods; Polyethylene Glycols; Rabbits - immunology; Radioimmunoassay; Serum Albumin, Bovine; Testis - analysis; Testosterone - analysis; Testosterone - blood; Tritium
• ISSN: 0022-4731
• DOI: 10.1016/0022-4731(73)90042-3

A radioimmunoassay technique, which is essentially a modification of the method described by Furuyama et al.[1], has been evaluated for the determination of testosterone in human peripheral plasma and rat testis tissue. The antiserum used was raised against testosterone-3-(0-carboxymethyl)-oxime-bovine serum albumin in female rabbits. It had an association constant of 5.5 × 10 9 1/mol, 4°C, at a dilution of 1 in 20,000. The procedure involved addition of [ 3H]-testosterone internal standard, extraction and chromatography of the plasma extracts on alumina micro-columns prior to assay. Testis tissue extracts were not chromatographed. Known amounts of standard testosterone were subjected to the same procedures. After incubation with antiserum for 16 h at 4°C total recovery from the extraction, chromatography (when used) and incubation procedures were measured in order to correct for losses. Either toluene scintillation fluid, dextran-coated charcoal or polyethylene glycol were used to separate free and bound testosterone. For human plasma as well as for testis tissue a good correlation was observed between results obtained with radioimmunoassay and a gas chromatographic method using electron capture detection of testosterone chloroacetate[12].