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Did we catch the point of the immunoassay principle correctly?

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Abstract

The present study was aimed at examining the degree of saturation of antibody in immunoassay. The results show that in equilibrium type immunoassay antibody is not fully occupied by antigen at any virtual point of the calibration curve since antibody saturation would lead to B/F=0. Calculations suggest that in an immunoassay meeting the condition p*→0 both relationships between antigen (p) and antibody (q) concentrations can be found (i.e. p<q; p>q). This is probably generally valid for any assay independently of the experimental technique and tracer used when a fixed amount of binder (antibody, receptor, etc.) is used for the analysis of a binding substance (antigen, ligand, etc.), and the proportion of their interaction is evaluated. Also, the appropriateness of the terms “saturation analysis” and “limited and/or excess reagent” assay for immunoassay is discussed.

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Abbreviations

p, p*, ps :

are the total concentrations of antigen, its labelled and unlabelled forms in the system (standard or unknown sample), respectively

q:

is the total concentration of antibody binding sites in the system

θ:

is the degree of antibody occupation by antigen

References

  1. J. C. TRAVIS, Fundamental of RIA and Other Ligand Assay. Anaheim, Scientific Newsletters Inc., 1977.

    Google Scholar 

  2. C. D. HAWKER, Anal. Chem., 45 (1973) 878A.

    Google Scholar 

  3. W. G. WOOD, G. SOKOLOWSKI, Radioimmunoassay in Theory and Practice. A Handbook for Laboratory Personnel. Konstanz, Schnetztor-Verlag, 1981.

    Google Scholar 

  4. L. SCHROTT, G. KLEINHAMMER, La Ricerca Clin. Lab., 8 (1978) 225.

    Google Scholar 

  5. R. P. EKINS, The future development of immunoassey, in: Radioimmunoassay and Related Procedures in Medicine, Vol. 1, IAEA, Vienna, 1978, p. 241.

  6. R. EKINS, Merits and disadvantages of different labels and methods of immunoassay, in: A. VOLLER, A. BARTLETT, D. BIDWELL (Eds), Immunoassay for 80s. MTP Press Limited, 1981, p. 5.

  7. R. P. EKINS, G. B. NEWMAN, J. L. H. O'RIORDAN, Theoretical aspects of “saturation” and radioimmunoassay, in: R. L. HAYES, F. A. GOSWITZ, B. E. P. MURPHY, Radioisotopes in Medicine: In Vitro Studies, Oak Ridge T. N., U.S. Atomic Energy Commision, 1968, p. 59.

    Google Scholar 

  8. R. EKINS, B. NEWMAN, Theoretical aspects of saturation analysis, in: E. DICZFALUSY, A. DICZFALUSY (Eds), Steroid Assay by Protein Binding. Karolinska Symposia on Research Methods in Reproductive Endocrinology. Acta Endocrinol., 64 (suppl. 147) (1970) 11.

  9. A. ZETTNER, Clin. Chem., 19 (1973) 699.

    PubMed  Google Scholar 

  10. J. RUŽICKA, J. STARÝ, Substoichiometry in Radiochemical Analysis. Pergamon Press Oxford, 1968.

    Google Scholar 

  11. S. A. BERSON, R. S. YALOW, Radioimmunoassay, in: S. A. BERSON, R. S. YALOW (Eds), Methods in Investigative and Diagnostic Endocrinology Vol. 2A, North-Holland Publishing Co., Amsterdam, 1973, p. 84.

    Google Scholar 

  12. D. S. SKELLEY, L. P. BROWN, P. K. BESCH, Clin. Chem. 19 (1973) 146.

    PubMed  Google Scholar 

  13. J. I. THORELL, S. M. LARSON, Radioimmunoassay and Related Techniques. Methodology and Clinical Applications, The C. V. Mosby Company, Saint Louis, 1978.

    Google Scholar 

  14. R. P. EKINS, in: K. E. KIRKHAM, W. M. HUNTER (Eds), Radioiummunoassay Methods, Churchill, Livingstone, 1971, p. 624.

    Google Scholar 

  15. D. RODBARD, in: Radioimmunoassay and Related Procedures in Medicine, Vol. 1, IAEA, Vienna, 1978, p. 269.

    Google Scholar 

  16. W. M. HUNTER, Recent Advances in Radioimmunoassay and Related Procedures, IAEA, Vienna, 1982, p. 3.

    Google Scholar 

  17. G. SCATCHARD, Ann. N. Y. Acad. Sci., 51 (1949) 660.

    Google Scholar 

  18. R. R. RUFFOLO, Jr., J. Auton. Pharmac., 2 (1982) 277.

    Google Scholar 

  19. J. S. WOODHEAD, G. M. ADDISON, C. N. HALES, J. L. H. O'RIORDAN, in: K. E. KIRKHAM, W. M. HUNTER (Eds), Radioimmunoassay Methods, Churchill, Livingstone, 1971, p. 467.

    Google Scholar 

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Authors and Affiliations

  1. Institute of Experimental Endocrinology, Centre of Physiological Sciences, Slovak Academy of Sciences, Bratislava, (Czechoslovakia)

    O. Földes

  2. Institute of Isotopes, Hungarian Academy of Sciences, Budapest, (Hungary)

    G. Tóth

Authors
  1. O. Földes
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  2. G. Tóth
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Földes, O., Tóth, G. Did we catch the point of the immunoassay principle correctly?. Journal of Radioanalytical and Nuclear Chemistry, Articles 120, 113–124 (1988). https://doi.org/10.1007/BF02037857

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  • DOI: https://doi.org/10.1007/BF02037857

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