Online citations, reference lists, and bibliographies.
← Back to Search

Enhancement Of Human Erythropoietin Production In Chinese Hamster Ovary Cells Through Supplementation Of 30Kc19-30Kc6 Fusion Protein

Hee Ho Park, Jeongseon Choi, Hong Jai Lee, Jina Ryu, J. Park, W. J. Rhee, T. Park
Published 2015 · Biology

Save to my Library
Download PDF
Analyze on Scholarcy
Abstract The importance of anti-apoptosis in mammalian cell culture has been widely recognized. We reported previously that expression of Bombyx mori 30 K genes in Chinese hamster ovary (CHO) cells increases recombinant protein production by inhibiting apoptosis and enhancing specific productivity. However, previous studies have shown expression of the anti-apoptotic 30Kc6 protein as inclusion bodies in Escherichia coli. 30Kc19 protein, another silkworm hemolymph protein, has cell-penetrating and recombinant protein productivity-improving properties, and we found that it improves soluble expression of its partner. In this study, we fused 30Kc6 with 30Kc19 as an expression partner to express a soluble fused protein and to deliver the protein to cells. Supplementing the recombinant 30Kc19-30Kc6 fusion protein in cell culture medium increased viability, effectively penetrated the cells, and inhibited CHO cell apoptosis by 68%. Moreover, the mitochondrial membrane potential and ATP generation also increased by 50% and 33%, respectively. Erythropoietin (EPO) productivity increased by > 30% because of the anti-apoptotic effect and increased specific productivity. These results demonstrate the potential use of this fusion protein as a supplement in mammalian cell culture during production of biopharmaceutical proteins.
This paper references
Purification of recombinant 30K protein produced in Escherichia coli and its anti-apoptotic effect in mammalian and insect cell systems
H. J. Park (2003)
A protein delivery system using 30Kc19 cell-penetrating protein originating from silkworm.
J. Park (2012)
Sodium butyrate inhibits histone deacetylation in cultured cells
E. P. Candido (1978)
Erythropoietin produced in a human cell line (Dynepo) has significant differences in glycosylation compared with erythropoietins produced in CHO cell lines.
Zahra Shahrokh (2011)
Production of recombinant protein therapeutics in cultivated mammalian cells
F. Wurm (2004)
Kinetic Effect of Silkworm Hemolymph on the Delayed Host Cell Death in an Insect Cell‐Baculovirus System
Won Jong Rhee (1999)
Beneficial effect of 30Kc6 gene expression on production of recombinant interferon-β in serum-free suspension culture of CHO cells
Tai Young Koo (2009)
Enhancement of recombinant human EPO production and glycosylation in serum-free suspension culture of CHO cells through expression and supplementation of 30Kc19
J. Park (2012)
N-acetylcysteine increases the biosynthesis of recombinant EPO in apoptotic Chinese hamster ovary cells.
K. H. Chang (1999)
Developmental and sex-dependent regulation of storage protein synthesis in the silkworm, Bombyx mori.
E. Mine (1983)
Silkworm hemolymph inhibits baculovirus-induced insect cell apoptosis.
W. J. Rhee (2000)
Inhibition of apoptosis by recombinant 30K protein originating from silkworm hemolymph.
E. J. Kim (2003)
The Growth Factor Inhibitor Suramin Reduces Apoptosis and Cell Aggregation in Protein‐Free CHO Cell Batch Cultures
J. A. Zanghi (2000)
Inhibition of Apoptosis by a Bombyx mori Gene
E. J. Kim (2004)
Selected amino acids protect hybridoma and CHO cells from elevated carbon dioxide and osmolality.
V. M. deZengotita (2002)
Enhancement of sialyltransferase-catalyzed transfer of sialic acid onto glycoprotein oligosaccharides using silkworm hemolymph and its 30K protein
S. Choi (2006)
Mammalian cell protein expression for biopharmaceutical production.
J. Zhu (2012)
Possible effect of 30K proteins in embryonic development of silkworm Bombyx mori.
B. Zhong (2005)
HIF-1α-dependent autophagy protects HeLa cells from fenretinide (4-HPR)-induced apoptosis in hypoxia.
X. Liu (2010)
The protection of hybridoma cells from apoptosis by caspase inhibition allows culture recovery when exposed to non-inducing conditions.
A. Tintó (2002)
Silkworm hemolymph as a substitute for fetal bovine serum in insect cell culture
S. Ha (1996)
Complete nucleotide sequences of major plasma protein genes of Bombyx mori.
S. Mori (1991)
Effect of N‐Acetylcystein on Butyrate‐Treated Chinese Hamster Ovary Cells To Improve the Production of Recombinant Human Interferon‐β‐1a
H. K. Oh (2005)
Expression of anti-apoptotic 30Kc6 gene inhibiting hyperosmotic pressure-induced apoptosis in antibody-producing Chinese hamster ovary cells
Zesong Wang (2012)
Expression of Bombyx mori 30Kc19 protein in Escherichia coli and its anti-apoptotic effect in Sf9 cell
W. J. Rhee (2009)
Beneficial effect of silkworm hemolymph on a CHO cell system: Inhibition of apoptosis and increase of EPO production.
S. Choi (2005)
The regulation of major haemolymph protein synthesis: changes in mRNA content during the development of Bombyx mori larvae
G. Bosquet (1989)
Enhancement of recombinant protein production in Chinese hamster ovary cells through anti‐apoptosis engineering using 30Kc6 gene
S. Choi (2006)
Anti-apoptosis engineering
Eun Jeong Kim (2003)
Enhancement of recombinant human EPO production and sialylation in chinese hamster ovary cells through Bombyx mori 30Kc19 gene expression
Zesong Wang (2011)
Efficient production of recombinant protein in Spodoptera frugiperda/AcNPV system utilizing silkworm hemolymph
S. Ha (2004)
Cellular engineering for the high-level production of recombinant proteins in mammalian cell systems
J. Park (2010)
Fusion protein linkers: property, design and functionality.
X. Chen (2013)
Recent advances in large-scale production of monoclonal antibodies and related proteins.
A. Shukla (2010)
Silkworm hemolymph as a potent inhibitor of apoptosis in Sf9 cells.
W. J. Rhee (2002)
Induction of G(2)/M phase arrest and apoptosis by oridonin in human laryngeal carcinoma cells.
Ning Kang (2010)
Enhancement of therapeutic monoclonal antibody production in CHO cells using 30Kc6 gene
Zesong Wang (2010)
Enhanced Human Thrombopoietin Production by Sodium Butyrate Addition to Serum‐Free Suspension Culture of Bcl‐2‐Overexpressing CHO Cells
Y. H. Sung (2005)
Cell engineering and cultivation of chinese hamster ovary (CHO) cells.
T. Omasa (2010)
Identification of MicroRNAs Involved in Hypoxia- and Serum Deprivation-Induced Apoptosis in Mesenchymal Stem Cells
Yu Nie (2011)
Overexpression of bcl-2 inhibits sodium butyrate-induced apoptosis in Chinese hamster ovary cells resulting in enhanced humanized antibody production.
N. Kim (2000)
Isolation and characterization of an apoptosis-inhibiting component from the hemolymph of Bombyx mori.
E. Kim (2001)
Understanding the mechanistic roles of 30Kc6 gene in apoptosis and specific productivity in antibody-producing Chinese hamster ovary cells
Zesong Wang (2012)
The genomic sequence of the Chinese hamster ovary (CHO)-K1 cell line
X. Xu (2011)
Identification and characterization of a novel cell-penetrating peptide of 30Kc19 protein derived from Bombyx mori
Hee Ho Park (2014)
Animal cell cultures: recent achievements and perspectives in the production of biopharmaceuticals
M. Butler (2005)
Inhibition of Human Cell Apoptosis by Silkworm Hemolymph
S. Choi (2002)
Stabilization of enzymes by the recombinant 30Kc19 protein
J. Park (2012)
CHO cells in biotechnology for production of recombinant proteins: current state and further potential
J. Y. Kim (2011)
Post-translational modifications of protein biopharmaceuticals.
G. Walsh (2010)

This paper is referenced by
Semantic Scholar Logo Some data provided by SemanticScholar