| DOI | Trouver le DOI : https://doi.org/10.1016/j.jbiotec.2016.04.028 |
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| Auteur | Rechercher : Delafosse, Laurence1; Rechercher : Xu, Ping1; Rechercher : Durocher, Yves1 |
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| Affiliation | - Conseil national de recherches Canada. Thérapeutique en santé humaine
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| Format | Texte, Article |
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| Sujet | Acylation; Polyplex; Transfection; Recombinant protein |
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| Résumé | Three commercially available linear polyethylenimines (25 kDa LPEI, 40 kDa PEI“Max” and PEIpro™) were compared regarding their potency to transfect serum-free growing and suspension-adapted HEK293 and CHO cells. We determined the optimal DNA:PEI ratios for maximal expression of the reporter gene SEAP while monitoring cytotoxicity following transfection. PEIs acylation was determined by ¹H NMR and their apparent size and polydispersity assessed by size-exclusion chromatography. The propensity of PEIs to condense plasmid DNA was evaluated by agarose-gel electrophoresis. The zeta potentials and particle sizes at optimal DNA:PEI ratio were analyzed. Polyplex attachment to the cells and internalization kinetics were monitored. The quantity of PEIpro™ needed to efficiently transfect the cells was significantly lower than with LPEI and PEI“Max” and, interestingly, the maximal amount of internalized PEIpro™-based polyplexes was approximately half of that observed with its counterparts. PEIpro™ was the largest and least polydisperse polymer, but also the most cytotoxic. The optimal transfection conditions were subsequently used to express three monoclonal antibodies at larger-scale. The use of the deacylated PEI“Max” and PEIpro™ resulted in a significant increase of recombinant protein expression compared to LPEI. These findings demonstrate the importance of properly choosing the most suitable polymers to obtain optimal recombinant protein transient expression. |
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| Date de publication | 2016-04-13 |
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| Maison d’édition | Elsevier |
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| Dans | |
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| Langue | anglais |
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| Publications évaluées par des pairs | Oui |
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| Identificateur | S0168165616302085 |
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| Numéro NPARC | 23000324 |
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| Exporter la notice | Exporter en format RIS |
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| Signaler une correction | Signaler une correction (s'ouvre dans un nouvel onglet) |
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| Identificateur de l’enregistrement | 2fd4c794-d8e7-4f43-b2ab-30dcfaadf161 |
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| Enregistrement créé | 2016-07-07 |
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| Enregistrement modifié | 2020-03-16 |
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