The kinetics of antibody binding to membrane antigens in solution and at the cell surface.
The reaction kinetics of 125I-labelled mouse monoclonal antibodies binding to three cell-surface antigens of rat thymocytes (Thy-1.1, W3/25) were studied. The differences between bivalent and univalent interactions were determined by using antibody in the F(ab')2 or Fab' form and by using antigen in polymeric or monomeric forms. Association rate constants (k+1), dissociation rate constants (k-1) and equilibrium constants were determined. Also, the dissociation kinetics of rabbit antibodies against rat Thy-1 antigen were studied. The major findings were as follows. (i) With F(ab')2 antibody there was no simple relationship between antigen density at the cell surface and extent of bivalent binding. Extensive univalent binding was observed unless the antibody had a high k-1 for the univalent interaction, in which case all binding was bivalent. (ii) k+1 values were similar for F(ab')2 or Fab' antibody, and for the different antibodies were in the range 0.8 x 10(5)--1.1 x 10(6) M-1.s-1. These differences were sufficient to affect the interpretation of serological assays with the different antibodies. (iii) Antibody bound bivalently dissociated much more slowly than that bound univalently. However, the k-1 values for the univalently bound antibody were sufficiently low in most cases that the lifetime of the univalent complex was similar to or greater than the time needed for the assay. Thus the results could be interpreted on the basis of irreversible reactions. The overall conclusion from the study is that for an understanding of the binding of antibody to cell-surface antigens the kinetics of the interaction are of major importance and theories based on equilibrium binding are inappropriate.
D000818:Animals; D000937:Antigen-Antibody Reactions; D000961:Antilymphocyte Serum / Q000276:immunology; D001853:Bone Marrow / Q000276:immunology; D001854:Bone Marrow Cells; D002462:Cell Membrane / Q000187:drug effects / Q000276:immunology; D002999:Clone Cells / Q000276:immunology; D003840:Deoxycholic Acid / Q000494:pharmacology; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D007700:Kinetics; D051379:Mice; D011817:Rabbits; D051381:Rats; D011946:Receptors, Antigen / Q000032:analysis; D013154:Spleen / Q000166:cytology / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology; D013696:Temperature; D013950:Thymus Gland / Q000166:cytology / Q000276:immunology
D000961:Antilymphocyte Serum; D007140:Immunoglobulin Fab Fragments; D011946:Receptors, Antigen; D003840:Deoxycholic Acid