4.7 Article

Poly(Beta-Amino Ester)s as High-Yield Transfection Reagents for Recombinant Protein Production

期刊

INTERNATIONAL JOURNAL OF NANOMEDICINE
卷 17, 期 -, 页码 4469-4479

出版社

DOVE MEDICAL PRESS LTD
DOI: 10.2147/IJN.S377371

关键词

non-viral gene transfer; transient transfection; mammalian cell expression systems; DNA delivery

资金

  1. NIH [R01EY031097, R01EB029455, R01CA228133, P41EB028239, R01CA240339]
  2. Emerson Collective Cancer Research Fund
  3. Bisciotti Foundation Translational Research Fund
  4. V Foundation Scholar Award
  5. National Science Foundation Graduate Research Fellowship Program award

向作者/读者索取更多资源

The study compared the use of PBAEs with linear 25 kDa PEI for transient transfection, showing that PBAEs demonstrated superior transfection efficiency and enhanced protein production in both HEK and CHO cells, making them potentially attractive reagents for recombinant protein production.
Purpose: Transient transfection is an essential tool for recombinant protein production, as it allows rapid screening for expression without stable integration of genetic material into a target cell genome. Poly(ethylenimine) (PEI) is the current gold standard for transient gene transfer, but transfection efficiency and the resulting protein yield are limited by the polymer's toxicity. This study investigated the use of a class of cationic polymers, poly(beta-amino ester)s (PBAEs), as reagents for transient transfection in comparison to linear 25 kDa PEI, a commonly used transfection reagent.Methods: Transfection efficiency and protein production were assessed in human embryonic kidney 293F (HEK) and Chinese hamster ovary-S (CHO) cell suspensions using PBAE-based nanoparticles in comparison to linear 25 kDa PEI. Production of both a cytosolic reporter and secreted antibodies was investigated. Results: In both HEK and CHO cells, several PBAEs demonstrated superior transfection efficiency and enhanced production of a cytosolic reporter compared to linear 25 kDa PEI. This result extended to secreted proteins, as a model PBAE increased the production of 3 different secreted antibodies compared to linear 25 kDa PEI at culture scales ranging from 20 to 2000 mL. In particular, non-viral gene transfer using the lead PBAE/plasmid DNA nanoparticle formulation led to robust transfection of mammalian cells across different constructs, doses, volumes, and cell types.Conclusion: These results show that PBAEs enhance transfection efficiency and increase protein yield compared to a widespread commercially available reagent, making them attractive candidates as reagents for use in recombinant protein production.

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