PMID: 8457598Mar 26, 1993Paper

Cooperativity in the antibody binding to surface-adsorbed antigen

Biochimica Et Biophysica Acta
M Werthén, H Nygren

Abstract

The binding to surface-adsorbed antigen of monoclonal mouse IgG-antibodies (mAbs), with two different affinities to dinitrophenyl (DNP), was measured by a calibrated ELISA. The concentration-dependence of antibody binding to surface-bound antigen of different epitope densities was analysed using Scatchard plots. The dissociation of bound tritium-labelled antibodies was measured in the presence of unlabelled antibodies in the bulk. At low surface concentration of bound anti-DNP, both high-affinity mAb and low-affinity mAb show a positive cooperativity in the binding reaction to antigen of high epitope density. Using antigen of lower epitope densities, the positive cooperativity is more pronounced for low-affinity clones. At higher surface concentrations of bound anti-DNP, the Scatchard plots indicate a negative cooperativity of binding, which is also implied by the increased dissociation found in the presence of antibodies in solution. The study confirms previous findings that the binding of antibodies to surface-bound antigen not only depends on intrinsic antibody affinity measured in solution. Other factors, such as self-interaction, also affect the heterogeneous binding reaction.

References

Jan 1, 1978·Methods in Enzymology·F W Dahlquist
Jun 12, 1985·Journal of Immunological Methods·H Nygren, M Stenberg
Nov 25, 1988·Journal of Immunological Methods·M Werthén, H Nygren
Aug 1, 1989·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·N S GreenspanL J Cooper
Aug 9, 1988·Journal of Immunological Methods·M StenbergH Nygren
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Jan 1, 1970·Advances in Clinical Chemistry·T Peters
Aug 3, 1984·Journal of Immunological Methods·G R NimmoM W Steward
Jan 1, 1982·Journal of Immunological Methods·C JacobsenJ Steensgaard

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Citations

Sep 27, 2002·Biophysical Journal·Simone KulinKristian Helmerson
Feb 11, 1999·European Journal of Pharmaceutical Sciences : Official Journal of the European Federation for Pharmaceutical Sciences·E E SiderisP E Macheras
Feb 1, 1997·Applied Biochemistry and Biotechnology·A Sadana, M Suturia
Jun 26, 2009·The Journal of Immunology : Official Journal of the American Association of Immunologists·Adam S ChervinDavid M Kranz

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