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Les Nanobodies, un nouvel outil de diagnostic de la maladie du court-noué de la vigne

Abstract : The grapevine fanleaf disease is mainly caused in Europe by the Grapevine fanleaf virus (GFLV) and the Arabis mosaic virus (ArMV). The principal mean to limit their spread, is to certify their absence in marketed grapevines by serological methods such as DAS-ELISA. Their unique biophysical and structural properties make the variable domains of heavy chain-only immunoglobulin, called Nanobodies (Nb) a real asset for the development of a diagnostic test against fanleaf disease viruses. I identified Nb able to detect a broad spectrum of viral isolates from two Nb collections against GFLV and ArMV. Their fusion to a fluorescent protein or to a bacterial alkaline phosphatase resulted in the production of efficient DAS-ELISA detection reagents. The atomic structure of a Nb/GFLV complex was solved at 2.8 Å by cryoelectron microscopy, allowing the precise mapping of the viral epitope. This result showed a maximum coverage of the viral particle by the Nb, leading to a maximal signal in DAS-ELISA. The full Nb tests against GFLV and ArMV were compared to commercial reagents and showed the superiority of the former in both sensitivity and specificity, opening the way for the development and commercialization of a new type of serological kits for the detection of grapevine viruses.
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Submitted on : Tuesday, June 26, 2018 - 2:56:44 PM
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  • HAL Id : tel-01823821, version 1

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Léa Ackerer. Les Nanobodies, un nouvel outil de diagnostic de la maladie du court-noué de la vigne. Immunologie. Université de Strasbourg, 2016. Français. ⟨NNT : 2016STRAJ103⟩. ⟨tel-01823821⟩

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