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Etude des interactions antigènes-anticorps par des techniques optiques : application aux protéines de surface erythrocytaires

Abstract : The objective of this work was double ; have a tool capable of characterizing antibodies in tenn of activity and to define the quality of the antigen-antibody interactions. The study of the antigens-antibody interactions was realized using optical techniques at the qualitative and quantitative and at the microscopie and macroscopic levels. The system erythrocyte antigens - monoclonal antibody anti-Glycophorin was chosen as an experimental model of specifie binding. We observed by microscopy in three dimensions the homogeneous distribution of the receptors on the cellular surface. The same technique was used to analyze the interactions between these receptors and the cytoske1eton. Our results showed interactions between Glycophorin C and Glycophorin B with the cytoskeleton and non interactions between Glycophorin A and the cytoskeleton. The quantification of the antigenic sites and the estimation of affinity constants of antibodies were carried out by flow cytometry using the Langmuir adsorption model. Our results are in accordance with previous studies. We also estimated, by means of the erythroaggregometer, the dissociation process of agglutinates subjected to constant or increasing shear rate in the gap of a Couette viscometer. This dissociation was followed by the measure of the backscattered light intensity by agg1utinates in suspension. Parameters characterizing erythrocyte agglutination were obtained and the specifie energy of dissociation was estimated from these parameters. Our work showed that the characterization of monoclonal antibodies can be carried out by the optical methods that we have developed.
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Natalia De Isla. Etude des interactions antigènes-anticorps par des techniques optiques : application aux protéines de surface erythrocytaires. Autre. Université Henri Poincaré - Nancy 1, 2003. Français. ⟨NNT : 2003NAN11308⟩. ⟨tel-01746925⟩



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