Journal
JOURNAL OF THEORETICAL BIOLOGY
Volume 290, Issue -, Pages 1-6Publisher
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
DOI: 10.1016/j.jtbi.2011.08.026
Keywords
Mathematical model; Neutrophils; Fc gamma RIIIb or CD16b receptors; Binding kinetics; Rate constants
Categories
Funding
- Siberian Branch of the Russian Academy of Science [2009-37, 2009-7]
- Presidium of the Russian Academy of Science [2009-27-15]
- Novosibirsk City Hall for young scientists [3210]
- Russian Ministry of Education and Science [P422, P1039, 14.740.11.0729, 14.740.11.1220, 14.740.11.1194, 14.740.11.0921]
- European Union [COST TD09/05]
- Grant Agency of Czech Republic [P302/10/1022]
- Academy of Sciences of the Czech Republic
- Ministry of Education Youth and Sports of the Czech Republic [LC535, LC06027, ME 919, AVOZ50040702, AVOZ50040507, COST-CZ LC11020]
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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 +/- 0.4) x 10(7) M(-1) min(-1), and the dissociation rate constant of (1.3 +/- 0.4) x 10(-1) min(-1). Based on the obtained values, the size of the receptor reaction site was estimated at approximately 1 nm. 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. (C) 2011 Elsevier. Ltd. All rights reserved.
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