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Master thesis

Medium optimization for the rhizosecretion of three monoclonal antibodies from Nicotiana tabacum hydroponic culture

Abstract : Plants are being developed as an inexpensive alternative platform for the production of recombinant pharmaceuticals, including monoclonal antibodies (mAbs). Transgenic plants were previously demonstrated to secrete the recombinant proteins from their roots in a process called rhizosecretion. Thus hydroponic cultivation of transgenic plants that allows harvesting of recombinant proteins from the hydroponic medium has been investigated for some years, but has not yet provided sufficient yields of target proteins for commercial viability. Scaling up the current in vitro production system could consist of the elaboration of an appropriate media for Nutrient Film Technique (NFT) ex vitro system. Medium supplemented with gelatin has previously shown 20 times-improvement of rhizosecreted antibody yields but is incompatible with NFT. In this project we have assessed loss in yields in the absence of gelatin of three monoclonal antibodies: M12 (anti-vitronectin), VRC01 (anti- HIV) and ?E559 (anti-rabies). Consistently, a 50% yield decrease occurred without gelatin, for each antibody. The NFT system has so far been recalcitrant to rhizosecretion with no detectable yields. Nitrate enriched hydroponic medium has previously been found to increase antibody secretion, by a mechanism currently thought to be osmolarity-related. A similar enrichment was applied to NFT medium in vitro in this project, and for the first time M12 antibody secretion could be detected and quantified. These results are encouraging for the scaling up of rhizosecretion to the NFT system as a recombinant protein production platform. Presently, plant yields remain dramatically lower than mammalian-cell production and plant molecular farming has not been embraced by pharmaceutical companies. However, such plant manufacturing systems could be of particular interest for the development of small scale, low-technology recombinant therapeutics production systems, highly needed to fight neglected diseases (e.g. rabies, tuberculosis) occurring in developing countries.
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Master thesis
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Submitted on : Wednesday, July 11, 2018 - 3:24:15 PM
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  • HAL Id : hal-01835626, version 1



Dorian Bocciarelli. Medium optimization for the rhizosecretion of three monoclonal antibodies from Nicotiana tabacum hydroponic culture. Environmental Sciences. 2014. ⟨hal-01835626⟩



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