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Développement d'une technologie hybride à base de microbilles pour la détection d'anticorps IgE : vers le diagnostic d'allergies

Abstract : Allergies represent a major social issue for modern societies. Current diagnostic tools enables to measure the concentration of specific IgE antibodies responsible for allergic response into the patient’s serum. The use of nano-micro particles decreases diffusion distances in order to shrink the size of device, reduce the length of time reaction and improve diagnostic efficiency. In this perspective, the PhD work develop an innovative method to detect IgE antibodies by using both 10µm polystyrene (PS) and 1µm superparamagnetic (SPM) microbeads.The PhD explores the idea of using both PS microbeads as support of immunoassay and magnetic properties of SPM microbeads in order to perform a magnetic sorting of PS microbeads. We have developped this technology in 3 different technological steps. First, biofunctionnalization is used to link allergens and antibodies onto the surface of microbeads intended to react with serum patient. Then, the magnetic sorting has been developped to isolate PS microbeads depending on the presence / absence of IgE antibodies onto their surface. The efficiency of such a process is proportional to IgE quantity per microbeads. Finally, a self-assembly process of microbeads called « gravitational assembly » has been developped by studying sedimentation and capillary phenomenon. It has been shown that this complementary technology to the capillary assembly technology extend the scope of assembly to dense system of colloidal suspension. I addition, this technology reaches a detection limit of 2 microbeads per ml.Clinical trials have been succesfully performed from serum patient allergic to peanut, hazelnut or shrimp. The process time, detection limit and dynamic range of the detection system have been studied and optimized. The proof of concept of IgE antibodies detection have been presented and points the way towards the multiplex diagnosis of allergies.
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Anthony Leonard. Développement d'une technologie hybride à base de microbilles pour la détection d'anticorps IgE : vers le diagnostic d'allergies. Biophysique [physics.bio-ph]. Université Grenoble Alpes, 2018. Français. ⟨NNT : 2018GREAY061⟩. ⟨tel-02053703⟩

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