Individual monoclonal antibodies vary greatly in the kinetics of binding to antigen (Mason and Williams, 1980; Ways and Parham, 1983; Parham, 1984a, b; Dower et al., 1984). The binding of individual monoclonal antibodies to the cell surface may reach saturation in as little as 15 min, or as long as 90 min (J. Goding, unpublished data). It seems clear that the rate of association is sometimes limited by more than simple diffusion. These results might be understood in terms of the need for an individual antigenic determinant that is recognized by a given monoclonal antibody to be in a particular (transient) conformation before the antibody can bind. Such slow 'on' rates might be expected to be highly dependent on temperature.
The highly individual kinetics of binding of monoclonal antibodies to their antigen makes it strongly advisable to keep incubation times constant from one experiment to another. Failure to do so may lead to errors in quantification and poor reproducibility in cases where the kinetics of binding are slow.
134 Monoclonal Antibodies: Principles and Practice
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