| DOI | Resolve DOI: https://doi.org/10.1016/j.jbiotec.2016.04.028 |
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| Author | Search for: Delafosse, Laurence1 ; Search for: Xu, Ping1 ; Search for: Durocher, Yves1 |
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| Name affiliation | - National Research Council Canada. Human Health Therapeutics
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| Format | Text |
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| Type | Article |
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| Journal title | Journal of Biotechnology |
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| ISSN | 0168-1656 |
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| Volume | 227 |
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| Pages | 103–111 |
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| Subject | Acylation; Polyplex; Transfection; Recombinant protein |
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| Abstract | 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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| Publication date | 2016-04-13 |
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| Publisher | Elsevier |
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| Terms of use | |
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| Language | English |
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| Peer reviewed | Yes |
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| Identifier | S0168165616302085 |
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| NPARC number | 23000324 |
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| Export citation | Export as RIS |
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| Report a correction | Report a correction |
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Record identifier | 2fd4c794-d8e7-4f43-b2ab-30dcfaadf161 | | Record created | 2016-07-07 |
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| Record modified | 2016-07-07 |
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