Increasing Transfection Efficiency of Lipoplexes by Modulating Complexation Solution for Transient Gene Expression

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 ca...

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Autores principales:	Jaemun Kim, Ji Yul Kim, Hyeonkyeong Kim, Eunsil Kim, Soonyong Park, Kyoung-Hwa Ryu, Eun Gyo Lee
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	transfection transient gene expression complexation solution CHO-S DC-Chol/DOPE Biology (General) QH301-705.5 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/ac35c97d0e7b4720943f3e6cc0bc4958
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spelling	oai:doaj.org-article:ac35c97d0e7b4720943f3e6cc0bc49582021-11-25T17:55:35ZIncreasing Transfection Efficiency of Lipoplexes by Modulating Complexation Solution for Transient Gene Expression10.3390/ijms2222123441422-00671661-6596https://doaj.org/article/ac35c97d0e7b4720943f3e6cc0bc49582021-11-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/22/12344https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067Transient 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 ζ 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.Jaemun KimJi Yul KimHyeonkyeong KimEunsil KimSoonyong ParkKyoung-Hwa RyuEun Gyo LeeMDPI AGarticletransfectiontransient gene expressioncomplexation solutionCHO-SDC-Chol/DOPEBiology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 12344, p 12344 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	transfection transient gene expression complexation solution CHO-S DC-Chol/DOPE Biology (General) QH301-705.5 Chemistry QD1-999
spellingShingle	transfection transient gene expression complexation solution CHO-S DC-Chol/DOPE Biology (General) QH301-705.5 Chemistry QD1-999 Jaemun Kim Ji Yul Kim Hyeonkyeong Kim Eunsil Kim Soonyong Park Kyoung-Hwa Ryu Eun Gyo Lee Increasing Transfection Efficiency of Lipoplexes by Modulating Complexation Solution for Transient Gene Expression
description	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 ζ 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.
format	article
author	Jaemun Kim Ji Yul Kim Hyeonkyeong Kim Eunsil Kim Soonyong Park Kyoung-Hwa Ryu Eun Gyo Lee
author_facet	Jaemun Kim Ji Yul Kim Hyeonkyeong Kim Eunsil Kim Soonyong Park Kyoung-Hwa Ryu Eun Gyo Lee
author_sort	Jaemun Kim
title	Increasing Transfection Efficiency of Lipoplexes by Modulating Complexation Solution for Transient Gene Expression
title_short	Increasing Transfection Efficiency of Lipoplexes by Modulating Complexation Solution for Transient Gene Expression
title_full	Increasing Transfection Efficiency of Lipoplexes by Modulating Complexation Solution for Transient Gene Expression
title_fullStr	Increasing Transfection Efficiency of Lipoplexes by Modulating Complexation Solution for Transient Gene Expression
title_full_unstemmed	Increasing Transfection Efficiency of Lipoplexes by Modulating Complexation Solution for Transient Gene Expression
title_sort	increasing transfection efficiency of lipoplexes by modulating complexation solution for transient gene expression
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/ac35c97d0e7b4720943f3e6cc0bc4958
work_keys_str_mv	AT jaemunkim increasingtransfectionefficiencyoflipoplexesbymodulatingcomplexationsolutionfortransientgeneexpression AT jiyulkim increasingtransfectionefficiencyoflipoplexesbymodulatingcomplexationsolutionfortransientgeneexpression AT hyeonkyeongkim increasingtransfectionefficiencyoflipoplexesbymodulatingcomplexationsolutionfortransientgeneexpression AT eunsilkim increasingtransfectionefficiencyoflipoplexesbymodulatingcomplexationsolutionfortransientgeneexpression AT soonyongpark increasingtransfectionefficiencyoflipoplexesbymodulatingcomplexationsolutionfortransientgeneexpression AT kyounghwaryu increasingtransfectionefficiencyoflipoplexesbymodulatingcomplexationsolutionfortransientgeneexpression AT eungyolee increasingtransfectionefficiencyoflipoplexesbymodulatingcomplexationsolutionfortransientgeneexpression
_version_	1718411822659272704

Increasing Transfection Efficiency of Lipoplexes by Modulating Complexation Solution for Transient Gene Expression

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