Immobilisation de biomolécules pour l’analyse multiparamétrique sur biopuces : application au génotypage érythrocytaire haut-débit

Abstract : The work reported in this thesis focuses on biomolecules immobilization for the development of multiparametric analysis tools on a biochip coupled with a colorimetric detection, applied to the characterization of biological samples and to diagnosis.The first concern was the development of high-throughput automated hybridization tests and immunotests on a filtration plate. This method led to the elaboration of an automated platform for extended blood group genotyping in collaboration with the Etablissement Français du Sang Rhône-Alpes (EFS-RA). It enables to analyze 96 samples in four hours and to characterize six genotypes per sample. Its analytical performances were validated on a panel of 293 blood donors.The second part of this work aimed to elaborate a new strategy for oligonucleotide immobilization on an innovative polymer (PolyshrinkTM) for the development of miniaturized analysis systems. Several approaches were evaluated and led to an in-situ immobilization of oligonucleotides in hydrogel dots technique. This method leads to 6 µm hydrogel dots with a diameter of 60 µm. Moreover it was demonstrated that such immobilized oligonucleotides were able to detect targets specifically and quantitatively using either a chemiluminescent or a colorimetric detection.
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Gaëlle Le Goff. Immobilisation de biomolécules pour l’analyse multiparamétrique sur biopuces : application au génotypage érythrocytaire haut-débit. Autre. Université Claude Bernard - Lyon I, 2011. Français. ⟨NNT : 2011LYO10196⟩. ⟨tel-00686770⟩

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