Valproic acid enhances recombinant mRNA and protein levels in transiently transfected Chinese hamster ovary cells.

Valproic acid (VPA) is a small molecule that inhibits histone deacetylase activity. Here we report that VPA increases recombinant mRNA and protein levels in transiently transfected CHO DG44 cells. In the presence of VPA, transient recombinant antibody yields of up to 40 mg/L were achieved in simple batch cultures. The steady-state levels of the IgG light and heavy chain mRNAs were nearly 10 times higher than in the untreated control transfection even though the level of transfected plasmid DNA was the same in the presence or absence of VPA. The combination of VPA treatment and incubation of the transfected cells in mildly hypothermic conditions resulted in recombinant antibody yields of over 90 mg/L by 6 days post-transfection in batch cultures. The results demonstrated that the treatment of transfected CHO DG44 cells with VPA is a cost-effective strategy for enhancing transient gene expression by increasing the transgene mRNA levels.

[1]  F. Wurm,et al.  Valproic acid: a viable alternative to sodium butyrate for enhancing protein expression in mammalian cell cultures. , 2008, Biotechnology and bioengineering.

[2]  A. Wieck,et al.  A review of valproate in psychiatric practice , 2009, Expert opinion on drug metabolism & toxicology.

[3]  Ping Zhu,et al.  The histone deacetylase inhibitor valproic acid selectively induces proteasomal degradation of HDAC2 , 2003, The EMBO journal.

[4]  F. Wurm,et al.  Rational vector design and multi-pathway modulation of HEK 293E cells yield recombinant antibody titers exceeding 1 g/l by transient transfection under serum-free conditions , 2008, Nucleic acids research.

[5]  C. Steidl,et al.  Results of a phase 2 study of valproic acid alone or in combination with all-trans retinoic acid in 75 patients with myelodysplastic syndrome and relapsed or refractory acute myeloid leukemia , 2005, Annals of Hematology.

[6]  A. El-Osta,et al.  Effect of valproic acid on radiation-induced DNA damage in euchromatic and heterochromatic compartments , 2008, Cell cycle.

[7]  P. Cheng,et al.  Activation of CMV promoter-controlled glycosyltransferase and β -galactosidase glycogenes by butyrate, tricostatin A, and 5-Aza-2′-deoxycytidine , 2005, Glycoconjugate Journal.

[8]  D. James,et al.  Control of Culture Environment for Improved Polyethylenimine‐Mediated Transient Production of Recombinant Monoclonal Antibodies by CHO Cells , 2006, Biotechnology progress.

[9]  Wenjun Guo,et al.  Induction of pluripotent stem cells by defined factors is greatly improved by small-molecule compounds , 2008, Nature Biotechnology.

[10]  N. Gattermann,et al.  Valproic acid for the treatment of myeloid malignancies , 2007, Cancer.

[11]  Sung Hyun Kim,et al.  Effect of Low Culture Temperature on Specific Productivity and Transcription Level of Anti‐4–1BB Antibody in Recombinant Chinese Hamster Ovary Cells , 2008, Biotechnology progress.

[12]  B. Howard,et al.  Expression of recombinant plasmids in mammalian cells is enhanced by sodium butyrate. , 1983, Nucleic acids research.

[13]  M. Bialer,et al.  Pharmacokinetics of valpromide after oral administration of a solution and a tablet to healthy volunteers , 2004, European Journal of Clinical Pharmacology.

[14]  Florian M. Wurm,et al.  Recombinant protein production by large-scale transient gene expression in mammalian cells: state of the art and future perspectives , 2007, Biotechnology Letters.

[15]  G. Lukács,et al.  Intracellular routing of plasmid DNA during non-viral gene transfer. , 2005, Advanced drug delivery reviews.

[16]  S. R. Fox,et al.  A detailed understanding of the enhanced hypothermic productivity of interferon‐γ by Chinese‐hamster ovary cells , 2005, Biotechnology and applied biochemistry.

[17]  M. Kay,et al.  Histone Modifications Are Associated with the Persistence or Silencing of Vector-mediated Transgene Expression in Vivo , 2022 .

[18]  F. Wurm,et al.  Mild Hypothermia Improves Transient Gene Expression Yields Several Fold in Chinese Hamster Ovary Cells , 2008, Biotechnology progress.

[19]  P. Chatellard,et al.  Transient gene expression: recombinant protein production with suspension-adapted HEK293-EBNA cells. , 2001, Biotechnology and bioengineering.

[20]  Jun-Feng Wang,et al.  Regulation of ER stress proteins by valproate: therapeutic implications. , 2002, Bipolar disorders.

[21]  H. Kohler,et al.  Histone deacetylases 1 and 2 act in concert to promote the G1-to-S progression. , 2010, Genes & development.

[22]  Gyun Min Lee,et al.  Effect of low culture temperature on specific productivity, transcription level, and heterogeneity of erythropoietin in Chinese hamster ovary cells. , 2003, Biotechnology and bioengineering.

[23]  F. Wurm,et al.  Serum‐free large‐scale transient transfection of CHO cells , 2004, Biotechnology and bioengineering.

[24]  Raymond Davis,et al.  Identification of novel small molecule enhancers of protein production by cultured mammalian cells , 2008, Biotechnology and bioengineering.

[25]  H. Koeffler,et al.  Histone Deacetylase Inhibitors Have a Profound Antigrowth Activity in Endometrial Cancer Cells , 2004, Clinical Cancer Research.

[26]  Amine Kamen,et al.  Large-Scale transfection of mammalian cells for the fast production of recombinant protein , 2006, Molecular biotechnology.

[27]  J. Cinatl,et al.  Sodium valproate inhibits in vivo growth of human neuroblastoma cells. , 1997, Anti-cancer drugs.

[28]  K. Struhl.  Histone acetylation and transcriptional regulatory mechanisms. , 1998, Genes & development.

[29]  Thomas D. Schmittgen,et al.  Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. , 2001, Methods.

[30]  F. Wurm,et al.  Scalable transient gene expression in Chinese hamster ovary cells in instrumented and non-instrumented cultivation systems , 2007, Biotechnology Letters.

[31]  Ping Zhu,et al.  Valproic acid defines a novel class of HDAC inhibitors inducing differentiation of transformed cells , 2001, The EMBO journal.

[32]  Zilong Shen,et al.  Valproic acid inhibits the growth of cervical cancer both in vitro and in vivo. , 2008, Journal of biochemistry.

[33]  J. Davie.  Inhibition of histone deacetylase activity by butyrate. , 2003, The Journal of nutrition.