Possible errors in the calculation of cell-binding parameters exemplified by the analysis of the IgG-Fc receptor system.

The binding parameters of monomeric and polymeric (immune complex with a molecular weight of 500,000 daltons) rabbit IgG to homologous Fc receptor-bearing alveolar macrophages were estimated, using corrected values for IgG and cell concentrations. Considering the maximum percentage of monomeric IgG binding to cells (2.7%) and the maximum percentage of cells binding monomeric IgG (32%) instead of the IgG and cell concentrations in the initial reaction mixture, a 36-fold increase of the equilibrium constant (K) (from 0.6 x 10(6) L/M to 21.3 x 10(6) L/M) and a 3-fold increase of the maximum number of IgG molecules able to bind to a single cell (n) (from 7.8 x 10(5) to 23.7 x 10(5] were registered. Since more than 60% of the polymeric IgG is bound to 46% of the macrophage population, the values of K (from 10.8 x 10(6) L/M to 15.6 x 10(6) L/M) and n (from 4.3 x 10(5) to 9.4 x 10(5] are only doubled by using the corrected values for IgG and cell concentrations. It results that the cytophilic fraction of the monomeric IgG, representing only 2.7% of total IgG, has a slightly higher affinity for the Fc receptors than the polymeric IgG. By considering the actual number of macrophages which bind IgG it is evident that the number of Fc receptors per cell is higher than that determined by the usual procedure which does not take into account cellular heterogeneity.