Abstract
Two progesterone-tyraminyl (125I) derivatives were prepared from progesterone-11-hemisuccinate and progesterone-3-0(carboxymethyl)oxime, respectively. The analytical performances of these tracers in the progesterone-anti progesterone-11-hemisuccinyl-BSA system were compared with those of tritiated radioligand assumed as a reference. For all of the 11 antisera tested much higher affinity and titers were observed in the case of ‘homologous’ 125I tracer, which however was found to involve drastic sensitivity losses. On the contrary, quite acceptable dose-response curves and affinity and titers close to those related to tritiated progesterone resulted with the heterologous tracer. These results, interpreted in terms of native ligand-tracer competitivity for the antibody sites, led to the choice of the C3-tyraminyl derivative for the development of a radioimmunoassay method to be compared with the reference tritiated progesterone systems. The two procedures, including minor differences in incubation conditions and B/F separation, were validated for the direct assay of unpurified dried serum extracts. In both cases, satifactory levels of sensitivity, precision and accuracy were proved, the estimates obtained being in a good agreement with data from the literature. On the basis of the present results, some general consideration on the optimization of steroid radioimmunoassay with radioiodinated tracers are tentatively drawn.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Abraham, G.E.: Radioimmunoassay of steroids in biological materials. Acta Endocrinol. (KbH.). 75, suppl. 183, 7 (1974)
Abraham, G.F., Swerdloff, R., Tulnhinsky, D., Odell, W.D.: Radioimmunoassay of plasma progesterone. J. Clin. Endocrin. Metab. 32, 619 (1971)
Cameron, E.H.D., Scarisbrick, J.J.: Radioimmunoassay of plasma progesterone. Clin. Chem. 19, 1403 (1973)
Cameron, E.H.D., Scarisbrick, J.J., Moleris, S.E., Hillier, S.G., Read, G.: 125I-iodohistamine derivatives as tracers for the radioimmunoassay of progestans. In: Steroid immunoassay, (H.E.D. Cameron, S.G. Hilleir, K. Griffiths, Eds.), p. 153. Cardiff: 1975 Alpha Omega Publ. Ltd.
Cameron, E.H.D., Scarisbrick, J.J., Morris, S.E., Hillier, S.G., Read, G.: Some aspects of the use of 125I-labelled ligands for steroid radioimmunoassay. J. Steroid Biochem 5, 749 (1974)
Comoglio, S., Saracco, B., Rosa, U.: Preparation of 17 β-oestradiol-6-(0-carboxymethyl)oxime (125I)-tyramine for oestradiol radioimmunoassay. J. Nucl. Biol. Med. 18, 98 (1974)
Crosignani, P.G., Malvano, R., Massaglia, A., Cozzani, P.: Problems connected with steroid radioimmunoassay. In: The endocrine function of the human testis, (V.H.T. James, M. Serio, L. Mantini, Eds.), p. 41. New York, London: Academic Press 1973
Diczfalusy, E.: A rapid 5 hour radioimmunoassay of progesterone and oestradiol in human plasma. Clin. Endocr. 4, 173 (1975)
Erlanger, B.F., Barck, F., Beiser, S.M., Lieberman, S.: Preparation and characterization of conjugates of bovine serum albumin with testosterone and with cortisone. J. Biol. Chemistry 228, 713 (1957)
Erlanger, B.F., Beiser, S.M., Barck, F., Edel, F., Lieberman, S.: The preparation of steroid-protein conjugates to elicit antihormonal antibodies. In: Methods in immunology and immunochemistry, (E.A. Williams, M.C. Chase, eds.), Vol. 1, p. 144, New York, London, Academic Press 1967
Hammond, G.L., Vinikka, L., Vihko, R.: Automation for some sex steroids with the use of both iodinated and tritiated ligands. Clin. Chem. 23, 1250 (1977)
Holmadahl, T.H., Johansson, E.D.B.: Peripheral plasma levels of 17-hydroxyprogesterone, progesterone and estradiol during normal menstrual cycles in women. Acta Endocrinol. (Kbh.) 71, 743 (1972)
Hunter, W.M., Grenwood, F.C.: Preparation of iodine-131 labelled human growth hormone of high specific activity. Nature 194, 495 (1962)
Jeffcoate, S.L., Gilby, E.D., Edwards, R.: The preparation and use of 125I steroid-albumin conjugates and tracers in steroid radioimmunoassay. Clin. Chim. Acta 43, 343 (1973)
Kutas, M., Chung, A., Bartos, D., Castro, A.: A simple proesterone radioimmunoassay without column chromatography. Steroid 20, 697 (1972)
Malvano, R., Rolleri E., Rosa, U.: Standardization and control of steroid radioimmunoassay in Radioimmunoassay and related procedures in medicine. Vol. II, p. 97. Vienna: IAEA 1974
Malvano, R., Zucchelli, G.C., Rosa, U., Salvetti, A.: Measurement of plasma renin activity by angiotensin I radioimmunoassay: (I) an assessment of some methodological aspects. J. Nucl. Biol. Med. 16, 24 (1972)
Nisonoff, A., Pressman, D.: Heterogeneity and average combining constant of antibodies from individual rabbits. J. Immun. 80, 417 (1958)
Niswender, G.D.: Influence of the site of conjugation on the specificity of antibodies to progesterone. Steroids 22, 413 (1973)
Niswender, G.D.: In: Steroid immunoassay, (H.E.D. Cameron, S.G. Hillier, K. Griffiths, Eds.), p. 165 (discussion). Cardiff: Alpha Omega Publ. Ltd. 1975
Scarisbrick, J.J., Cameron, E.H.D.: Radioimmunoassay of progesterone: comparison of [1,2,6,7-3H4]-Progesterone and Progesterone-[125I]-iodohistamine radioligands. J. Steroid Biochem. 6, 51 (1975)
Scarisbrick, J.J., Read, G., Cameron, E.H.D.: Radioimmunoassay of progesterone: comparison of 3H- and 125I-labelled radioligands. J. Endocrinol. 61, xli-xlii (1974)
Wars, P.W., Hunter, W.M.: A method for labelling oestradiol-17 with radioiodine for radioimmunoassay. J. Endocrin. 57, xlvii (1973)
Youssefneiadian, E., Forensa, E., Collins, W.P., Sommerville, I.F.: Radioimmunoassay of plasma progesterone. J. Steroid Biochem. 3, 893 (1972)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Malvano, R., Zucchelli, G.C., Galbiati, A. et al. Radioimmunoassay of progesterone in crude serum extracts using tritiated and (125I) radioiodinated tracers. Eur J Nucl Med 4, 301–308 (1979). https://doi.org/10.1007/BF00304889
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF00304889