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Distribution function approach to the study of the kinetics of IgM antibody binding to Fc\(\gamma\)RIIIb (CD16b) receptors on neutrophils by flow cytometry. (English) Zbl 1397.92295
Summary: Though flow cytometry provides the entire distribution of cellular fluorescence (i.e., “fluorescence profile”), only mean fluorescence data are usually considered in studies of ligand-receptor binding. In this study, we presented a method of the treatment of the temporal evolution of the whole fluorescence profile with a comprehensive statistical approach extended to the reversible binding case. The method was demonstrated in the study of the 1D3 IgM monoclonal antibodies binding to Fc\(\gamma\)RIIIb receptors (CD16b) on neutrophils. Kinetic experiments were carried out using a FACSCalibur (Becton Dickinson, USA) flow cytometer. For each of four donors, we obtained the distribution of the number of Fc\(\gamma\)RIIIb surface receptors for neutrophils and the rate constants per receptor: the association rate constant of \((2.7\pm 0.4)\times 10^7\mathrm M^{-1}\min^{-1}\), and the dissociation rate constant of \((1.3\pm 0.4)\times 10^{-1}\min^{-1}\). Based on the obtained values, the size of the receptor reaction site was estimated at approximately 1nm. It was found, that cell receptors distributions differed sufficiently between donors in mean and the skewness values, whereas the coefficient of variation (i.e., the ratio of the standard deviation to the mean) did not vary significantly.
MSC:
92C45 Kinetics in biochemical problems (pharmacokinetics, enzyme kinetics, etc.)
92C55 Biomedical imaging and signal processing
62P10 Applications of statistics to biology and medical sciences; meta analysis
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