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Solid-Phase Radioimmunoassay of Steroids

  • Stephen A. Tillson
  • Ian H. Thorneycroft
  • Guy E. Abraham
  • Rex J. Scaramuzzi
  • Burton V. Caldwell

Abstract

There have been several procedures reported which offer the new investigator interested in developing a radioimmunoassay for steroid hormones a choice of methods for separating the free from bound steroid. Midgley et al. (1969) have described a means for accomplishing separation, based on precipitation of bound antibody-steroid with a second antibody. This separation usually requires incubation for several days. Mikhail et al. (1970) separate free from bound estradiol by polymerizing the antibody, but this adds a high-speed centrifugation step. The recently described solid-phase system for estradiol (Abraham, 1969; Abraham and Odell, 1970) would appear to offer many advantages over the above procedures, particularly the unique feature of accomplishing the equilibration and the separation of free from bound steroid in a single step. It will be the principal aim of this paper to describe in detail the several essential criteria which must be examined for each antiserum, central to its use in the development of new methods for the radioimmunoassay of Steroids. Chemical, immunologic, and endocrinologic aspects of steroid-protein conjugates have been described by Lieberman et al. (1959). Pressman and Grossberg (1968, 1970) have discussed chemical and structural concepts which are involved in antibody-antigen interactions. However, the effects of certain physicochemical factors, such as ionic strength, hydrogen ion concentration, and temperature, on potential radioimmunoassay systems for progesterone and testosterone have not been previously examined in detail and will be described below as they apply to the solid-phase assay of these hormones.

Keywords

Normal Rabbit Serum Immunologic Method Percent Serum Polystyrene Tube Free Hormone 
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References

  1. Abraham, G. E. 1969. Solid-phase radioimmunoassay of estradiol-17β. J. Clin. Endocr., 29:866–870.PubMedCrossRefGoogle Scholar
  2. -and W. D. Odell. Solid-phase radioimmunoassay of serum estradiol-17β: A semi-automated approach. 1970. In Péron, F. G., and B. V. Caldwell, eds. Immunologic Methods in Steroid Determination. New York, Appleton-Century-Crofts.Google Scholar
  3. Catt, K. J. 1969. In Margaulies, M., ed. Protein and Polypeptide Hormones. New York, Excerpta Medica Foundation, Part 3, pp. 639–642.Google Scholar
  4. -and G. W. Tregear. 1967. Solid-phase radioimmunoassay in antibody-coated tubes. Science, 158:1570–1572.PubMedCrossRefGoogle Scholar
  5. Ganguly, M., and U. Westphal. 1968. Steroid-protein interactions. XVII. Influence of solvent environment on interaction between α1 acid glycoproteins and progesterone. J. Biol. Chem., 243:6130–6139.PubMedGoogle Scholar
  6. --- R. H. Carneghan, and U. Westphal. 1967. Steroid-protein interactions. XIV. Interaction between human α1 acid glycoprotein and progesterone. Biochemistry (Wash.), 6:2803–2814.CrossRefGoogle Scholar
  7. Greenwood, F. C., W. M. Hunter, and J. S. Glover. 1963. The preparations of I131-labeled human growth hormone of high specific radioactivity. Biochem. J., 89:114–123.PubMedGoogle Scholar
  8. Grossberg, A. L., C. C. Chen, L. Rendina, and D. Pressman. 1962. Specific cation effects with antibody to a hapten with a positive charge. J. Immun., 88:600–603.PubMedGoogle Scholar
  9. Horton, R., T. Kato, and R. Sherins. 1967. A rapid method for the estimation of testosterone in male plasma. Steroids, 10:245–256.PubMedCrossRefGoogle Scholar
  10. Jiang, N. S., and R. J. Ryan. 1969. Radioimmunoassay for estrogens: A preliminary communication. Mayo Clin. Proc., 44:461–465.PubMedGoogle Scholar
  11. Kato, T., and R. Horton. 1968. A rapid method for the estimation of testosterone in female plasma. Steroids, 12:631–650.PubMedCrossRefGoogle Scholar
  12. Korenman, S. G. 1968. Radio-ligand binding assay of specific estrogens using a soluble uterine macromolecule. J. Clin. Endocr., 28:127–130.PubMedCrossRefGoogle Scholar
  13. Lieberman, S., B. F. Erlanger, S. M. Beiser, and F. J. Agate, Jr. 1959. Steroid-protein conjugates: Their chemical immunochemical, and endocrinology properties. Recent Progr. Hormone Res., 15:165–200.Google Scholar
  14. Maeda, R., M. Okamoto, L. C. Wegienka, and P. H. Forsham. 1969. A clinically useful method for plasma testosterone determination. Steroids, 13:83–99.PubMedCrossRefGoogle Scholar
  15. Mayes, D., and C. A. Nugent. 1968. Determination of plasma testosterone by the use of competitive protein binding. J. Clin. Endocr., 28:1169–1176.PubMedCrossRefGoogle Scholar
  16. Midgley, A. R., Jr., G. D. Niswender, and J. S. Ram. 1969. Hapten-radioimmunoassay: A general procedure for the estimation of standard and other haptenic substances. Steroids, 13:731–737.PubMedCrossRefGoogle Scholar
  17. Mikhail, G., C. Wu, M. Ferin, and R. L. Vande Wiele. 1970. Radioimmunoassay of plasma estrogens: Use of polymerized antibodies. In Péron, F. G., and Caldwell, B. V., eds. Immunologic Methods in Steroid Determination. New York, Appleton-Century-Crofts.Google Scholar
  18. Murphy, B. E. P. 1967. Some studies of the protein-binding steroids and their application to the routine micro and ultramicro measurements of various steroids in body fluids by competitive protein-binding radioassay. J. Clin. Endocr., 27:973–990.PubMedCrossRefGoogle Scholar
  19. Neill, J. D., E. D. B. Johnansson, J. K. Datta, and E. Knobil. 1967. Relationship between the plasma levels of luteinizing hormone and progesterone during the normal menstrual cycle. J. Clin. Endocr., 27:1167–1173.PubMedCrossRefGoogle Scholar
  20. Pressman, D., and A. L. Grossberg. 1968. The Structural Basis of Antibody Specificity. New York, W. A. Benjamin, Inc.Google Scholar
  21. --- and A. L. Grossberg. 1970. Structural basis of antibody specificity. In Péron, F. G., and Caldwell, B. V., eds. Immunologic Methods in Steroid Determination. New York, Appleton-Century-Crofts.Google Scholar
  22. --- A. Nisonoff, and G. Radzimski. 1961a. Specific anion effects with antibenzoate antibody. J. Immun., 86:35–41.PubMedGoogle Scholar
  23. --- A. Nisonoff, G. Radzimski, and A. Shaw. 1961b. Nature of the active site of antibenzoate antibodies: Further evidence for the presence of tyrosine. J. Immun., 86:489–495.PubMedGoogle Scholar
  24. Rosenfield, R. L., W. R. Eberlein, and A. M. Bongiovanni. 1969. Measurement of plasma testosterone by means of competitive protein binding analysis. J. Clin. Endocr., 29:854–859.PubMedCrossRefGoogle Scholar
  25. Sandberg, A. A., H. Rosenthal, S. L. Schneider, and W. R. Slaunwhite, Jr. 1966. Protein-steroid interactions and their role in the transport and metabolism of steroids. In Pincus, G., T. Nakao, and J. F. Tait, eds. Steroid Dynamics. New York, Academic Press, Inc., pp. 1–62.Google Scholar
  26. Thorneycroft, I. H., S. A. Tillson, G. E. Abraham, R. J. Scaramuzzi, and B. V. Caldwell. 1970. Preparation and purification of antibodies to steroids. In Péron, F. G., and Caldwell, B. V., eds. Immunologic Methods in Steroid Determination. New York, Appleton-Century-Crofts.Google Scholar
  27. Yoshimi, T., and M. B. Lipsett. 1968. The measurement of plasma progesterone. Steroids, 11:527–540.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1970

Authors and Affiliations

  • Stephen A. Tillson
    • 1
  • Ian H. Thorneycroft
    • 1
  • Guy E. Abraham
    • 1
  • Rex J. Scaramuzzi
    • 1
  • Burton V. Caldwell
    • 1
  1. 1.Worcester Foundation for Experimental BiologyShrewsburyUSA

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