Comparative study of polyethylenimines for transient gene expression in mammalian HEK293 and CHO cells

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DOIResolve DOI: http://doi.org/10.1016/j.jbiotec.2016.04.028
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TypeArticle
Journal titleJournal of Biotechnology
ISSN0168-1656
Volume227
Pages103111
SubjectAcylation; Polyplex; Transfection; Recombinant protein
AbstractThree 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.
Publication date
PublisherElsevier
LanguageEnglish
AffiliationHuman Health Therapeutics; National Research Council Canada
Peer reviewedYes
IdentifierS0168165616302085
NPARC number23000324
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Record identifier2fd4c794-d8e7-4f43-b2ab-30dcfaadf161
Record created2016-07-07
Record modified2016-07-07
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