Literature DB >> 22415736

Effect of addition of 'carrier' DNA during transient protein expression in suspension CHO culture.

Ketaki Pradhan1, Mugdha Gadgil.   

Abstract

Transient protein expression using polyethyleneimine as a transfection agent is useful for the rapid production of small amounts of recombinant proteins. It is known that an increase in extracellular DNA concentration during transfection can lead to a nonlinear increase in intracellular DNA concentration. We present an approach that hypothesizes that this nonlinearity can be used to decrease the amount of plasmid required for productive transfections. Through addition of non coding 'carrier' DNA to increase total DNA concentration during transfection, we report a statistically significant increase in protein (IgG) expression per unit plasmid used for transfection. This approach could be useful to increase protein yields for large scale transfections under conditions where plasmid availability is limited.

Entities:  

Year:  2012        PMID: 22415736      PMCID: PMC3488370          DOI: 10.1007/s10616-012-9435-4

Source DB:  PubMed          Journal:  Cytotechnology        ISSN: 0920-9069            Impact factor:   2.058


  27 in total

1.  Quantitative analysis of correlation between number of nuclear plasmids and gene expression activity after transfection with cationic liposomes.

Authors:  Rieko Tachibana; Hideyoshi Harashima; Naoko Ide; Sachiko Ukitsu; Yasuko Ohta; Norio Suzuki; Hiroshi Kikuchi; Yasuo Shinohara; Hiroshi Kiwada
Journal:  Pharm Res       Date:  2002-04       Impact factor: 4.200

2.  Reproducible transfection in the presence of carrier DNA using FuGENE6 and Lipofectamine2000.

Authors:  Takao Susa; Takako Kato; Yukio Kato
Journal:  Mol Biol Rep       Date:  2007-05-17       Impact factor: 2.316

3.  100-liter transient transfection.

Authors:  Philippe Girard; Madiha Derouazi; Gwendoline Baumgartner; Michaela Bourgeois; Martin Jordan; Barbara Jacko; Florian M Wurm
Journal:  Cytotechnology       Date:  2002-01       Impact factor: 2.058

4.  Mechanistic studies on aggregation of polyethylenimine-DNA complexes and its prevention.

Authors:  Vikas K Sharma; Mini Thomas; Alexander M Klibanov
Journal:  Biotechnol Bioeng       Date:  2005-06-05       Impact factor: 4.530

Review 5.  Large-scale transfection of mammalian cells for the fast production of recombinant protein.

Authors:  Phuong Lan Pham; Amine Kamen; Yves Durocher
Journal:  Mol Biotechnol       Date:  2006-10       Impact factor: 2.695

6.  Design of Experiment in CHO and HEK transient transfection condition optimization.

Authors:  Frederic Bollin; Vincent Dechavanne; Laurent Chevalet
Journal:  Protein Expr Purif       Date:  2011-02-24       Impact factor: 1.650

7.  Serum-free large-scale transient transfection of CHO cells.

Authors:  Madiha Derouazi; Philippe Girard; Frédéric Van Tilborgh; Keyvan Iglesias; Natalie Muller; Martin Bertschinger; Florian M Wurm
Journal:  Biotechnol Bioeng       Date:  2004-08-20       Impact factor: 4.530

8.  The efficiency of nuclear plasmid DNA delivery is a critical determinant of transgene expression at the single cell level.

Authors:  Dominic J Glover; Denisse L Leyton; Gregory W Moseley; David A Jans
Journal:  J Gene Med       Date:  2010-01       Impact factor: 4.565

9.  SEAP expression in transiently transfected mammalian cells grown in serum-free suspension culture.

Authors:  Ernst-Jürgen Schlaeger; Eric A Kitas; Arnulf Dorn
Journal:  Cytotechnology       Date:  2003-05       Impact factor: 2.058

10.  Transient gene expression in mammalian cells grown in serum-free suspension culture.

Authors:  E J Schlaeger; K Christensen
Journal:  Cytotechnology       Date:  1999-07       Impact factor: 2.058

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  4 in total

1.  A Novel Terminator Primer and Enhancer Reagents for Direct Expression of PCR-Amplified Genes in Mammalian Cells.

Authors:  Mikiko Nakamura; Ayako Suzuki; Junko Akada; Tohru Yarimizu; Ryo Iwakiri; Hisashi Hoshida; Rinji Akada
Journal:  Mol Biotechnol       Date:  2015-08       Impact factor: 2.695

2.  A simple, rapid method for evaluation of transfection efficiency based on fluorescent dye.

Authors:  Lin Peng; Wendian Xiong; Yanfei Cai; Yun Chen; Yang He; Jianfeng Yang; Jian Jin; Huazhong Li
Journal:  Bioengineered       Date:  2016-09-27       Impact factor: 3.269

3.  HP1α targets the chromosomal passenger complex for activation at heterochromatin before mitotic entry.

Authors:  Jan G Ruppert; Kumiko Samejima; Melpomeni Platani; Oscar Molina; Hiroshi Kimura; A Arockia Jeyaprakash; Shinya Ohta; William C Earnshaw
Journal:  EMBO J       Date:  2018-02-21       Impact factor: 14.012

4.  Modulating CRISPR-Cas Genome Editing Using Guide-Complementary DNA Oligonucleotides.

Authors:  Thomas Swartjes; Peng Shang; Dennis T M van den Berg; Tim Künne; Niels Geijsen; Stan J J Brouns; John van der Oost; Raymond H J Staals; Richard A Notebaart
Journal:  CRISPR J       Date:  2022-07-18
  4 in total

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