Regular Article
Antibody Affinities and Relative Titers in Polyclonal Populations: Surface Plasmon Resonance Analysis of anti-DNA Antibodies

https://doi.org/10.1006/abbi.1999.1469Get rights and content

Abstract

This paper presents the equations and methodology for the measurement and interpretation of apparent dissociation constants for polyclonal populations of antibodies, where antigen is kept trace relative to antibody concentration. Surface plasmon resonance is used to determine Kds for the binding of anti-DNA antibodies to trace amounts of DNA antigen on a chip. Since the approach taken relies on equilibrium measurements, kinetic mass transport artifacts are avoided. The apparent Kd is a weighted average of all the Kds for the clonally related subpopulations within the polyclonal pool, where each weighting factor is the relative titer (fractional presence) of the subpopulation. Titration curves appear as if there is one monoclonal population with that titer-weighted-average Kd. Implications of changes in the antibody affinity distribution within the population are discussed. The equations described herein provide a better physical understanding of the apparent Kd that is obtained when a heterogeneous population of receptors is titrated against a trace ligand.

References (27)

  • T.P. Werblin et al.

    Immunochemistry

    (1972)
  • J. Piehler et al.

    J. Immunol. Methods

    (1997)
  • R.W. Glaser

    Anal. Biochem.

    (1993)
  • L. Nieba et al.

    Anal. Biochem.

    (1996)
  • B. Liedberg et al.

    Sensors Actuators

    (1983)
  • M. Malmqvist

    Curr. Opin. Immunol.

    (1993)
  • M.R. Bowles et al.

    Anal. Biochem.

    (1997)
  • H.N. Eisen et al.

    Biochemistry

    (1964)
  • M. Heidelberger et al.

    J. Exp. Med.

    (1935)
  • K. Landsteiner et al.

    J. Exp. Med.

    (1936)
  • A. Nisonoff et al.

    J. Immunol.

    (1958)
  • L. Pauling et al.

    J. Am. Chem. Soc.

    (1944)
  • S.H. Huang et al.

    Pharm. Rev.

    (1982)
  • Cited by (0)

    1

    To whom correspondence should be addressed. Fax: (619) 546-2307. E-mail: [email protected].

    View full text