Increasing transfection efficiency of lipoplexes by modulating complexation solution for transient gene expression

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Title
Increasing transfection efficiency of lipoplexes by modulating complexation solution for transient gene expression
Author(s)
Jaemun Kim; Ji Yul Kim; Hyeonkyeong KimEunsil KimSoonyong ParkKyoung Hwa RyuEun Gyo Lee
Bibliographic Citation
International Journal of Molecular Sciences, vol. 22, no. 22, pp. 12344-12344
Publication Year
2021
Abstract
Transient gene expression is a suitable tool for the production of biopharmaceutical candidates in the early stage of development and provides a simple and rapid alternative to the generation of stable cell line. In this study, an efficient transient gene expression methodology using DC-Chol/DOPE cationic liposomes and pDNA in Chinese hamster ovary suspension cells was established through screening of diverse lipoplex formation conditions. We modulated properties of both the liposome formation and pDNA solution, together called complexation solutions. Protein expression and cellular cytotoxicity were evaluated following transfection over the cell cultivation period to select the optimal complexation solution. Changes in hydrodynamic size, polydispersity index, and potential of the liposomes and lipoplexes were analyzed depending on the various pH ranges of the complexation solutions using dynamic light scattering. The transfer of lipoplexes to the cytosol and their conformation were traced using fluorescence analysis until the early period of transfection. As a result, up to 1785 mg/L and 191 mg/L of human Fc protein and immunoglobulin G (bevacizumab), respectively, were successfully produced using acidic liposome formation and alkaline pDNA solutions. We expect that this lipoplex formation in acidic and alkaline complexation solutions could be an effective methodology for a promising gene delivery strategy.
Keyword
TransfectionTransient gene expressionComplexation solutionCHO-SDC-Chol/DOPE
ISSN
1661-6596
Publisher
MDPI
DOI
http://dx.doi.org/10.3390/ijms222212344
Type
Article
Appears in Collections:
Division of Bio Technology Innovation > BioProcess Engineering Center > 1. Journal Articles
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