Optimisation de la production de protéines recombinantes avec des cellules de tabac en suspension
Bibliographic record
Abstract
RESUME La demande croissante pour des proteines recombinantes a faible cout stimule l’industrie a developper de nouvelles plateformes de production. Parmi celles-ci, la plateforme vegetale in vitro gagne en interet. Permettant l’utilisation d’un milieu de culture entierement synthetique, exempt de proteines et qui ne favorise pas le developpement de pathogenes humains, les cellules de plantes peuvent effectuer des modifications post-traductionnelles et mener a des molecules bioactives. Plusieurs proteines produites en plantes sont d’ailleurs en phase d'essais cliniques. Toutefois, les faibles rendements en proteines recombinantes obtenus avec les cellules de plantes limitent la competitivite de cette plateforme novatrice. Les travaux de recherche regroupes dans cette these visent l’amelioration des rendements en proteines recombinantes produites par des cellules de tabac en suspension. L'objectif principal a consiste en l’optimisation de la production de proteines recombinantes en cellules de tabac, dans le contexte d'un bioprocede en bioreacteur. En objectifs specifiques, la co-culture de cellules de plantes avec Agrobacterium permettant d'exprimer de facon transitoire un gene d’interet et un suppresseur viral, ainsi que l’identification et l’isolation de promoteurs forts inductibles au froid ont ete etudiees. De maniere a presenter une vue globale de la situation actuelle du marche et de l’industrie, une revue detaillee a ete realisee et soumise au journal Plant Biotechnology Journal (fevrier 2010). Dans la premiere partie de cet article, l'ensemble des plateformes de production de proteines recombinantes etablies de meme que celles en emergence sont decrites en detail et leurs niches specifiques sont presentees. Par la suite, les innovations recentes rapportees dans la litterature pour la plateforme vegetale in vitro, telles que l’optimisation des cassettes d’expression et des parametres de bioprocedes et l’utilisation de suppresseurs viraux de repression genique sont presentees et discutees.----------ABSTRACT The continuously increasing demand for bioactive recombinant proteins at low cost stimulates the industry to develop new production platforms. Among them, in vitro plant biotechnology is gaining interest. Allowing the use of a fully synthetic protein-free medium that do not promote the development of human pathogens, plant cells can perform post-translational modifications and lead to bioactive therapeutics. Various human proteins produced in plant cell cultures are currently under clinical trials. However, low recombinant protein yields obtained with plant cells limit the competitiveness of this new platform. The research in this thesis is aimed at raising recombinant protein yields from tobacco cells in suspensions. The main objective was to optimize recombinant protein production in tobacco cells in a bioreactor process. Secondary objectives were to study plant cell coculture with Agrobacterium to transiently express a gene of interest and a viral suppressor and to identify and isolate strong cold-inducible promoters. In order to present an overview of the actual situation in the industry, an exhaustive literature review was produced and submitted to the Plant Biotechnology Journal (February 2010). In the first part of this review manuscript, all the established recombinant protein production platforms as well as the main emerging platforms are described in detail with their specific applications. Upcoming innovations for the in vitro plant biotechnology platform such as the optimization of expression cassettes, of bioprocess parameters and the use of viral suppressors are then presented and discussed.
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How this classification was reachedexpand
Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.001 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.001 | 0.001 |
| Insufficient payload (model declined to judge) | 0.000 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one teacher head, not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".