Cancer Immunology, Immunotherapy

, Volume 54, Issue 6, pp 513–516

Binding parameters of antibodies: pseudo-affinity and other misconceptions

Position Paper

Abstract

In order to evaluate and compare monoclonal antibodies (Abs), functional affinities are generally determined. While the equations that define affinity relate to monovalent interactions, it has been considered that the binding of Abs to multivalent antigens such as the cell surface could be described by an apparent, or functional affinity. We demonstrate here that this concept is incorrect, since the binding interactions that occur cannot be described in terms of a functional affinity, and the values that are obtained serve only to obscure the true interactions. Bivalent Ab binding must be considered to be an irreversible reaction, in most cases. A correct understanding of Ab binding will be useful in the further development of Abs for therapeutic purposes.

Keywords

Antibody therapy Antibody affinity Antibody binding parameters 

References

  1. 1.
    Berzofsky JA, Epstein SL, Berkower IJ (1989) Antigen-antibody interactions and monoclonal antibodies. In: Paul WE (ed) Fundamental immunology, 2 edn. Raven Press, New York, p 325Google Scholar
  2. 2.
    Crothers DM, Metzger H (1972) The influence of polyvalency on the binding properties of antibodies. Immunochemistry 9:341CrossRefGoogle Scholar
  3. 3.
    Dower SK, Ozato K, Segal DM (1984) The interaction of monoclonal antibodies with MHC class I antigens on mouse spleen cells. I: analysis of the mechanism of binding. J Immunol 132:751Google Scholar
  4. 4.
    Finkelstein MS, Uhr JW (1966) Antibody formation. V: the avidity of γM and γG guinea pig antibodies to bacteriophage Φ X174. J Immunol 97:565Google Scholar
  5. 5.
    Genentech Inc (2003) Herceptin U.S. prescribing informationGoogle Scholar
  6. 6.
    Hanna R, Ong GL, Mattes MJ (1996) Processing of antibodies bound to B-cell lymphomas and other hematological malignancies. Cancer Res 56:3062PubMedGoogle Scholar
  7. 7.
    Hauck ML, Dewhirst MW, Zalutsky MR (1996) The effects of clinically relevant hyperthermic temperatures on the kinetic binding parameters of a monoclonal antibody. Nucl Med Biol 23:551CrossRefGoogle Scholar
  8. 8.
    Hornick CL, Karush F (1972) Antibody affinity. III: the role of multivalence. Immunochemistry 9:325CrossRefGoogle Scholar
  9. 9.
    IDEC Pharmaceutical Corp (2002) Rituxin U.S. prescribing informationGoogle Scholar
  10. 10.
    Karush F (1976) Multivalent binding and functional affinity. Comtemp Top Mol Immunol 5:217Google Scholar
  11. 11.
    Karush F (1978) The affinity of antibody: range, variability, and the role of multivalence. In: Litman G, Good RA (eds) Comprehensive Immunology, vol 5, Immunoglobulins. Plenum Press, New YorkGoogle Scholar
  12. 12.
    Kaufman EN, Jain RK (1992) Effect of bivalent interaction upon apparent antibody affinity: experimental confirmation of theory using fluorescence photobleaching and implications for antibody binding assays. Cancer Res 52:4157Google Scholar
  13. 13.
    Klotz IM (1982) Numbers of receptor sites from Scatchard graphs: facts and fantasies. Science 217:1247Google Scholar
  14. 14.
    Kohler G, Milstein C (1975) Continuous cultures of fused cells secreting antibody of defined specificity. Nature 256:495PubMedGoogle Scholar
  15. 15.
    Kyriakos RJ, Shih LB, Ong GL, Patel K, Goldenberg DM, Mattes MJ (1992) The fate of antibodies bound to the surface of tumor cells in vitro. Cancer Res 52:835Google Scholar
  16. 16.
    Lindmo T, Boven E, Cuttitta F, Fedorko J, Bunn PA (1984) Determination of the immunoreactive fraction of radiolabeled monoclonal antibodies by linear extrapolation to binding at infinite antigen excess. J Immunol Methods 72:77CrossRefPubMedGoogle Scholar
  17. 17.
    Mason DW, Williams AF (1980) The kinetics of antibody binding to membrane antigens in solution and at the cell surface. Biochem J 187:1Google Scholar
  18. 18.
    Mason DW, Williams AF (1986) Kinetics of antibody reactions and the analysis of cell surface antigens. In: Weir DM (ed) Handbook of experimental immunology, 4th edn. Blackwell Scientific, Oxford, p 38.1Google Scholar
  19. 19.
    Mattes MJ (1995) Limitations of the Lindmo method in determining antibody immunoreactivity. Int J Cancer 61:286Google Scholar
  20. 20.
    Mattes MJ (1997) Binding parameters of antibodies reacting with multivalent antigens: functional affinity or pseudo-affinity. J Immunol Methods 202:97CrossRefGoogle Scholar
  21. 21.
    Mattes MJ, Griffiths GL, Diril H, Goldenberg DM, Ong GL, Shih LB (1994) Processing of antibody-radioisotope conjugates after binding to the surface of tumor cells. Cancer 73(Suppl):787Google Scholar
  22. 22.
    Nygren H, Czerkinsky C, Stenberg M (1985) Dissociation of antibodies bound to surface-immobilized antigen. J Immunol Methods 85:87CrossRefGoogle Scholar
  23. 23.
    Ong GL, Mattes MJ (1993) Re-evaluation of the concept of functional affinity as applied to bivalent antibody binding to cell surfaces. Mol Immunol 30:1455CrossRefGoogle Scholar
  24. 24.
    Parham P, Brodsky FM (1981) Partial purification and some properties of BB7.2. A cytotoxic monoclonal antibody with specificity for HLA-A2 and a variant of HLA-A28. Hum Immunol 3:227CrossRefGoogle Scholar
  25. 25.
    Reynolds JA (1979) Interaction of divalent antibody binding to cell surface antigens. Biochemistry 18:264Google Scholar
  26. 26.
    Scatchard G (1949) The attractions of proteins for small molecules and ions. Ann NY Acad Sci 51:660Google Scholar
  27. 27.
    Trucco M, de Petris S (1981) Determination of equilibrium binding parameters of monoclonal antibodies specific for cell surface antigens. In: Lefkovits I, Pernis B (eds) Immunological Methods, vol II. Academic Press, New York, p 1Google Scholar
  28. 28.
    Ways JP, Parham P (1983) The binding of monoclonal antibodies to cell surface molecules. Biochem J 216:423Google Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  1. 1.Center for Molecular Medicine and ImmunologyBellevilleUSA

Personalised recommendations