Applied Microbiology and Biotechnology

, Volume 93, Issue 3, pp 917–930 | Cite as

CHO cells in biotechnology for production of recombinant proteins: current state and further potential

  • Jee Yon Kim
  • Yeon-Gu Kim
  • Gyun Min Lee


Recombinant Chinese hamster ovary cells (rCHO) cells have been the most commonly used mammalian host for large-scale commercial production of therapeutic proteins. Recent advances in cell culture technology for rCHO cells have achieved significant improvement in protein production leading to titer of more than 10 g/L to meet the huge demand from market needs. This achievement is associated with progression in the establishment of high and stable producer and the optimization of culture process including media development. In this review article, we focus on current strategies and achievements in cell line development, mainly in vector engineering and cell engineering, for high and stable protein production in rCHO cells. The approaches that manipulate various DNA elements for gene targeting by site-specific integration and cis-acting elements to augment and stabilize gene expression are reviewed here. The genetic modulation strategy by “direct” cell engineering with growth-promoting and/or productivity-enhancing factors and omics-based approaches involved in transcriptomics, proteomics, and metabolomics to pursue cell engineering are also presented.


CHO cells Cell line development Vector engineering Cell engineering Omics-based approaches 



This research was supported in part by the World Class University program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (MEST, R31-2008-000-10071-0), the Converging Research Center Program through the NRF funded by the MEST (2009–0082276), and a grant from the Fundamental R&D Program for Technology of World Premier Materials funded by the Ministry of Knowledge Economy, Republic of Korea.


  1. Adams JM, Cory S (2001) Life-or-death decisions by the Bcl-2 protein family. Trends Biochem Sci 26:61–66Google Scholar
  2. Arden N, Betenbaugh MJ (2004) Life and death in mammalian cell culture: strategies for apoptosis inhibition. Trends Biotechnol 22:174–180Google Scholar
  3. Arden N, Majors BS, Ahn SH, Oyler G, Betenbaugh MJ (2007) Inhibiting the apoptosis pathway using MDM2 in mammalian cell cultures. Biotechnol Bioeng 97:601–614Google Scholar
  4. Astley K, Al-Rubeai M (2008) The role of Bcl-2 and its combined effect with p21CIP1 in adaptation of CHO cells to suspension and protein-free culture. Appl Microbiol Biotechnol 78:391–399Google Scholar
  5. Baik JY, Lee GM (2010) A DIGE approach for the assessment of differential expression of the CHO proteome under sodium butyrate addition: Effect of Bcl-xL overexpression. Biotechnol Bioeng 105:358–367Google Scholar
  6. Baik JY, Lee MS, An SR, Yoon SK, Joo EJ, Kim YH, Park HW, Lee GM (2006) Initial transcriptome and proteome analyses of low culture temperature-induced expression in CHO cells producing erythropoietin. Biotechnol Bioeng 93:361–371Google Scholar
  7. Baik JY, Joo EJ, Kim YH, Lee GM (2008) Limitations to the comparative proteomic analysis of thrombopoietin producing Chinese hamster ovary cells treated with sodium butyrate. J Biotechnol 133:461–468Google Scholar
  8. Baik JY, Ha TK, Kim YH, Lee GM (2011) Proteomic understanding of intracellular responses of recombinant Chinese hamster ovary cells adapted to grow in serum-free suspension culture. Biotechnol Prog. doi: 10.1002/btpr.685
  9. Barnes LM, Bentley CM, Dickson AJ (2003) Stability of protein production from recombinant mammalian cells. Biotechnol Bioeng 81:631–639Google Scholar
  10. Barron N, Kumar N, Sanchez N, Doolan P, Clarke C, Meleady P, O'Sullivan F, Clynes M (2011) Engineering CHO cell growth and recombinant protein productivity by overexpression of miR-7. J Biotechnol 151:204–211Google Scholar
  11. Becker E, Florin L, Pfizenmaier K, Kaufmann H (2008) An XBP-1 dependent bottle-neck in production of IgG subtype antibodies in chemically defined serum-free Chinese hamster ovary (CHO) fed-batch processes. J Biotechnol 135:217–223Google Scholar
  12. Benton T, Chen T, McEntee M, Fox B, King D, Crombie R, Thomas TC, Bebbington C (2002) The use of UCOE vectors in combination with a preadapted serum free, suspension cell line allows for rapid production of large quantities of protein. Cytotechnology 38:43–46Google Scholar
  13. Bi JX, Shuttleworth J, Al-Rubeai M (2004) Uncoupling of cell growth and proliferation results in enhancement of productivity in p21CIP1-arrested CHO cells. Biotechnol Bioeng 85:741–749Google Scholar
  14. Borth N, Mattanovich D, Kunert R, Katinger H (2005) Effect of increased expression of protein disulfide isomerase and heavy chain binding protein on antibody secretion in a recombinant CHO cell line. Biotechnol Prog 21:106–111Google Scholar
  15. Bradley SA, Ouyang A, Purdie J, Smitka TA, Wang T, Kaerner A (2010) Fermentanomics: monitoring mammalian cell cultures with NMR spectroscopy. J Am Chem Soc 132:9531–9533Google Scholar
  16. Cabaniols JP, Ouvry C, Lamamy V, Fery I, Craplet ML, Moulharat N, Guenin SP, Bedut S, Nosjean O, Ferry G, Devavry S, Jacqmarcq C, Lebuhotel C, Mathis L, Delenda C, Boutin JA, Duchâteau P, Cogé F, Pâques F (2010) Meganuclease-driven targeted integration in CHO-K1 cells for the fast generation of HTS-compatible cell-based assays. J Biomol Screen 15:956–967Google Scholar
  17. Carlage T, Hincapie M, Zang L, Lyubarskaya Y, Madden H, Mhatre R, Hancock WS (2009) Proteomic profiling of a high-producing Chinese hamster ovary cell culture. Anal Chem 81:7357–7362Google Scholar
  18. Carvalhal AV, Marcelino I, Carrondo MJ (2003) Metabolic changes during cell growth inhibition by p27 overexpression. Appl Microbiol Biotechnol 63:164–173Google Scholar
  19. Chiang GG, Sisk WP (2005) Bcl-xL mediates increased production of humanized monoclonal antibodies in Chinese hamster ovary cells. Biotechnol Bioeng 91:779–792Google Scholar
  20. Choi SS, Rhee WJ, Kim EJ, Park TH (2006) Enhancement of recombinant protein production in Chinese hamster ovary cells through anti-apoptosis engineering using 30Kc6 gene. Biotechnol Bioeng 95:459–467Google Scholar
  21. Chong WP, Goh LT, Reddy SG, Yusufi FN, Lee DY, Wong NS, Heng CK, Yap MG, Ho YS (2009) Metabolomics profiling of extracellular metabolites in recombinant Chinese hamster ovary fed-batch culture. Rapid Commun Mass Spectrom 23:3763–3771Google Scholar
  22. Chong WP, Reddy SG, Yusufi FN, Lee DY, Wong NS, Heng CK, Yap MG, Ho YS (2010) Metabolomics-driven approach for the improvement of Chinese hamster ovary cell growth: overexpression of malate dehydrogenase II. J Biotechnol 147:116–121Google Scholar
  23. Chong WP, Yusufi FN, Lee DY, Reddy SG, Wong NS, Heng CK, Yap MG, Ho YS (2011) Metabolomics-based identification of apoptosis-inducing metabolites in recombinant fed-batch CHO culture media. J Biotechnol 151:218–224Google Scholar
  24. Chung JY, Lim SW, Hong YJ, Hwang SO, Lee GM (2004) Effect of doxycycline-regulated calnexin and calreticulin expression on specific thrombopoietin productivity of recombinant Chinese hamster ovary cells. Biotechnol Bioeng 85:539–546Google Scholar
  25. Cost GJ, Freyvert Y, Vafiadis A, Santiago Y, Miller JC, Rebar E, Collingwood TN, Snowden A, Gregory PD (2010) BAK and BAX deletion using zinc-finger nucleases yields apoptosis-resistant CHO cells. Biotechnol Bioeng 105:330–340Google Scholar
  26. Crea F, Sarti D, Falciani F, Al-Rubeai M (2006) Over-expression of hTERT in CHO K1 results in decreased apoptosis and reduced serum dependency. J Biotechnol 121:109–123Google Scholar
  27. Davis R, Schooley K, Rasmussen B, Thomas J, Reddy P (2000) Effect of PDI overexpression on recombinant protein secretion in CHO cells. Biotechnol Prog 16:736–743Google Scholar
  28. De Leon Gatti M, Wlaschin KF, Nissom PM, Yap M, Hu WS (2007) Comparative transcriptional analysis of mouse hybridoma and recombinant Chinese hamster ovary cells undergoing butyrate treatment. J Biosci Bioeng 103:82–91Google Scholar
  29. Doolan P, Melville M, Gammell P, Sinacore M, Meleady P, McCarthy K, Francullo L, Leonard M, Charlebois T, Clynes M (2008) Transcriptional profiling of gene expression changes in a PACE-transfected CHO DUKX cell line secreting high levels of rhBMP-2. Mol Biotechnol 39:187–199Google Scholar
  30. Doolan P, Meleady P, Barron N, Henry M, Gallagher R, Gammell P, Melville M, Sinacore M, McCarthy K, Leonard M, Charlebois T, Clynes M (2010) Microarray and proteomics expression profiling identifies several candidates, including the valosin-containing protein (VCP), involved in regulating high cellular growth rate in production CHO cell lines. Biotechnol Bioeng 106:42–56Google Scholar
  31. Dreesen IA, Fussenegger M (2011) Ectopic expression of human mTOR increases viability, robustness, cell size, proliferation, and antibody production of Chinese hamster ovary cells. Biotechnol Bioeng 108:853–866Google Scholar
  32. Figueroa B Jr, Ailor E, Osborne D, Hardwick JM, Reff M, Betenbaugh MJ (2007) Enhanced cell culture performance using inducible anti-apoptotic genes E1B-19K and Aven in the production of a monoclonal antibody with Chinese hamster ovary cells. Biotechnol Bioeng 97:877–892Google Scholar
  33. Florin L, Pegel A, Becker E, Hausser A, Olayioye MA, Kaufmann H (2009) Heterologous expression of the lipid transfer protein CERT increases therapeutic protein productivity of mammalian cells. J Biotechnol 141:84–90Google Scholar
  34. Fussenegger M, Mazur X, Bailey JE (1997) A novel cytostatic process enhances the productivity of Chinese hamster ovary cells. Biotechnol Bioeng 55:927–939Google Scholar
  35. Fussenegger M, Schlatter S, Dätwyler D, Mazur X, Bailey JE (1998) Controlled proliferation by multigene metabolic engineering enhances the productivity of Chinese hamster ovary cells. Nat Biotechnol 16:468–472Google Scholar
  36. Gammell P, Barron N, Kumar N, Clynes M (2007) Initial identification of low temperature and culture stage induction of miRNA expression in suspension CHO-K1 cells. J Biotechnol 130:213–218Google Scholar
  37. Girod PA, Zahn-Zabal M, Mermod N (2005) Use of the chicken lysozyme 5′ matrix attachment region to generate high producer CHO cell lines. Biotechnol Bioeng 91:1–11Google Scholar
  38. Girod PA, Nguyen DQ, Calabrese D, Puttini S, Grandjean M, Martinet D, Regamey A, Saugy D, Beckmann JS, Bucher P, Mermod N (2007) Genome-wide prediction of matrix attachment regions that increase gene expression in mammalian cells. Nat Methods 4:747–753Google Scholar
  39. Goudar C, Biener R, Boisart C, Heidemann R, Piret J, de Graaf A, Konstantinov K (2010) Metabolic flux analysis of CHO cells in perfusion culture by metabolite balancing and 2D [13C, 1H] COSY NMR spectroscopy. Metab Eng 12:138–149Google Scholar
  40. Gupta P, Lee KH (2007) Genomics and proteomics in process development: opportunities and challenges. Trends Biotechnol 25:324–330Google Scholar
  41. Hacker DL, De Jesus M, Wurm FM (2009) 25 years of recombinant proteins from reactor-grown cells—where do we go from here? Biotechnol Adv 27:1023–1027Google Scholar
  42. Han YK, Kim YG, Kim JY, Lee GM (2010) Hyperosmotic stress induces autophagy and apoptosis in recombinant Chinese hamster ovary cell culture. Biotechnol Bioeng 105:1187–1192Google Scholar
  43. Han YK, Ha TK, Lee SJ, Lee JS, Lee GM (2011) Autophagy and apoptosis of recombinant Chinese hamster ovary cells during fed-batch culture: effect of nutrient supplementation. Biotechnol Bioeng 108:2182–2192Google Scholar
  44. Hayduk EJ, Lee KH (2005) Cytochalasin D can improve heterologous protein productivity in adherent Chinese hamster ovary cells. Biotechnol Bioeng 90:354–364Google Scholar
  45. Hayes NV, Smales CM, Klappa P (2010) Protein disulfide isomerase does not control recombinant IgG4 productivity in mammalian cell lines. Biotechnol Bioeng 105:770–779Google Scholar
  46. Huang Y, Li Y, Wang YG, Gu X, Wang Y, Shen BF (2007) An efficient and targeted gene integration system for high-level antibody expression. J Immunol Methods 322:28–39Google Scholar
  47. Hwang SO, Lee GM (2008) Nutrient deprivation induces autophagy as well as apoptosis in Chinese hamster ovary cell culture. Biotechnol Bioeng 99:678–685Google Scholar
  48. Hwang SO, Lee GM (2009) Effect of Akt overexpression on programmed cell death in antibody-producing Chinese hamster ovary cells. J Biotechnol 139:89–94Google Scholar
  49. Hwang SO, Chung JY, Lee GM (2003) Effect of doxycycline-regulated ERp57 expression on specific thrombopoietin productivity of recombinant CHO cells. Biotechnol Prog 19:179–184Google Scholar
  50. Jaluria P, Betenbaugh M, Konstantopoulos K, Shiloach J (2007) Enhancement of cell proliferation in various mammalian cell lines by gene insertion of a cyclin-dependent kinase homolog. BMC Biotechnol 7:e71Google Scholar
  51. Jayapal KP, Wlaschin KF, Hu WS, Yap MG (2007) Recombinant protein therapeutics from CHO cells—20 years and counting. Chem Eng Prog 103:40–47Google Scholar
  52. Kameyama Y, Kawabe Y, Ito A, Kamihira M (2010) An accumulative site-specific gene integration system using Cre recombinase-mediated cassette exchange. Biotechnol Bioeng 105:1106–1114Google Scholar
  53. Kantardjieff A, Nissom PM, Chuah SH, Yusufi F, Jacob NM, Mulukutla BC, Yap M, Hu WS (2009) Developing genomic platforms for Chinese hamster ovary cells. Biotechnol Adv 27:1028–1035Google Scholar
  54. Kantardjieff A, Jacob NM, Yee JC, Epstein E, Kok YJ, Philp R, Betenbaugh M, Hu WS (2010) Transcriptome and proteome analysis of Chinese hamster ovary cells under low temperature and butyrate treatment. J Biotechnol 145:143–159Google Scholar
  55. Kaufmann H, Mazur X, Fussenegger M, Bailey JE (1999) Influence of low temperature on productivity, proteome and protein phosphorylation of CHO cells. Biotechnol Bioeng 63:573–582Google Scholar
  56. Kennard ML, Goosney DL, Monteith D, Zhang L, Moffat M, Fischer D, Mott J (2009) The generation of stable, high MAb expressing CHO cell lines based on the artificial chromosome expression (ACE) technology. Biotechnol Bioeng 104:540–553Google Scholar
  57. Kim NS, Lee GM (2001) Overexpression of bcl-2 inhibits sodium butyrate-induced apoptosis in Chinese hamster ovary cells resulting in enhanced humanized antibody production. Biotechnol Bioeng 71:184–193Google Scholar
  58. Kim NS, Lee GM (2002a) Inhibition of sodium butyrate-induced apoptosis in recombinant Chinese hamster ovary cells by constitutively expressing antisense RNA of caspase-3. Biotechnol Bioeng 78:217–228Google Scholar
  59. Kim NS, Lee GM (2002b) Response of recombinant Chinese hamster ovary cells to hyperosmotic pressure: effect of Bcl-2 overexpression. J Biotechnol 95:237–248Google Scholar
  60. Kim SH, Lee GM (2007a) Down-regulation of lactate dehydrogenase-A by siRNAs for reduced lactic acid formation of Chinese hamster ovary cells producing thrombopoietin. Appl Microbiol Biotechnol 74:152–159Google Scholar
  61. Kim SH, Lee GM (2007b) Functional expression of human pyruvate carboxylase for reduced lactic acid formation of Chinese hamster ovary cells (DG44). Appl Microbiol Biotechnol 76:659–665Google Scholar
  62. Kim MS, Lee GM (2008) Use of Flp-mediated cassette exchange in the development of a CHO cell line stably producing erythropoietin. J Microbiol Biotechnol 18:1342–1351Google Scholar
  63. Kim YG, Lee GM (2009) Bcl-xL overexpression does not enhance specific erythropoietin productivity of recombinant CHO cells grown at 33 °C and 37 °C. Biotechnol Prog 25:252–256Google Scholar
  64. Kim JM, Kim JS, Park DH, Kang HS, Yoon J, Baek K, Yoon Y (2004) Improved recombinant gene expression in CHO cells using matrix attachment regions. J Biotechnol 107:95–105Google Scholar
  65. Kim JD, Yoon Y, Hwang HY, Park JS, Yu S, Lee J, Baek K, Yoon J (2005) Efficient selection of stable chinese hamster ovary (CHO) cell lines for expression of recombinant proteins by using human interferon beta SAR element. Biotechnol Prog 21:933–937Google Scholar
  66. Kim MS, Kim WH, Lee GM (2007) A simple analysis system for the estimation of recombination efficiency using fluorescence-activated cell sorting. J Biotechnol 127:373–384Google Scholar
  67. Kim MS, Kim WH, Lee GM (2008) Characterization of site-specific recombination mediated by Cre recombinase during the development of erythropoietin producing CHO cell lines. Biotechnol Bioprocess Eng 13:418–423Google Scholar
  68. Kim YG, Kim JY, Lee GM (2009a) Effect of XIAP overexpression on sodium butyrate-induced apoptosis in recombinant Chinese hamster ovary cells producing erythropoietin. J Biotechnol 144:299–303Google Scholar
  69. Kim YG, Kim JY, Mohan C, Lee GM (2009b) Effect of Bcl-xL overexpression on apoptosis and autophagy in recombinant Chinese hamster ovary cells under nutrient-deprived condition. Biotechnol Bioeng 103:757–766Google Scholar
  70. Kim JY, Kim YG, Han YK, Choi HS, Kim YH, Lee GM (2011) Proteomic understanding of intracellular responses of recombinant Chinese hamster ovary cells cultivated in serum-free medium supplemented with hydrolysates. Appl Microbiol Biotechnol 89:1917–1928Google Scholar
  71. Kito M, Itami S, Fukano Y, Yamana K, Shibui T (2002) Construction of engineered CHO strains for high-level production of recombinant proteins. Appl Microbiol Biotechnol 60:442–448Google Scholar
  72. Koh TC, Lee YY, Chang SQ, Nissom PM (2009) Identification and expression analysis of miRNAs during batch culture of HEK-293 cells. J Biotechnol 140:149–155Google Scholar
  73. Ku SC, Ng DT, Yap MG, Chao SH (2008) Effects of overexpression of X-box binding protein 1 on recombinant protein production in Chinese hamster ovary and NS0 myeloma cells. Biotechnol Bioeng 99:155–164Google Scholar
  74. Kumar N, Gammell P, Meleady P, Henry M, Clynes M (2008) Differential protein expression following low temperature culture of suspension CHO-K1 cells. BMC Biotechnol 8:e42Google Scholar
  75. Kuystermans D, Al-Rubeai M (2009) cMyc increases cell number through uncoupling of cell division from cell size in CHO cells. BMC Biotechnol 9:e76Google Scholar
  76. Kuystermans D, Dunn MJ, Al-Rubeai M (2010) A proteomic study of cMyc improvement of CHO culture. BMC Biotechnol 10:e25Google Scholar
  77. Lao MS, Toth D (1997) Effects of ammonium and lactate on growth and metabolism of a recombinant Chinese hamster ovary cell culture. Biotechnol Prog 13:688–691Google Scholar
  78. Lee SK, Lee GM (2003) Development of apoptosis-resistant dihydrofolate reductase efficient Chinese hamster ovary cell line. Biotechnol Bioeng 82:872–876Google Scholar
  79. Lee KH, Harrington MG, Bailey JE (1996a) Two-dimensional electrophoresis of proteins as a tool in the metabolic engineering of cell cycle regulation. Biotechnol Bioeng 50:336–340Google Scholar
  80. Lee KH, Sburlati A, Renner WA, Bailey JE (1996b) Deregulated expression of cloned transcription factor E2F-1 in Chinese hamster ovary cells shifts protein patterns and activates growth in protein-free medium. Biotechnol Bioeng 50:273–279Google Scholar
  81. Lee MS, Kim KW, Kim YH, Lee GM (2003) Proteome analysis of antibody-expressing CHO cells in response to hyperosmotic pressure. Biotechnol Prog 19:1734–1741Google Scholar
  82. Lee YY, Wong KT, Tan J, Toh PC, Mao Y, Brusic V, Yap MG (2009) Overexpression of heat shock proteins (HSPs) in CHO cells for extended culture viability and improved recombinant protein production. J Biotechnol 143:34–43Google Scholar
  83. Levine B, Klionsky DJ (2004) Development by self-digestion: molecular mechanisms and biological functions of autophagy. Dev Cell 6:463–477Google Scholar
  84. Li J, Huang Z, Sun X, Yang P, Zhang Y (2006) Understanding the enhanced effect of dimethyl sulfoxide on hepatitis B surface antigen expression in the culture of Chinese hamster ovary cells on the basis of proteome analysis. Enzyme Microb Technol 38:372–380Google Scholar
  85. Majors BS, Arden N, Oyler GA, Chiang GG, Pederson NE, Betenbaugh MJ (2008) E2F-1 overexpression increases viable cell density in batch cultures of Chinese hamster ovary cells. J Biotechnol 138:103–106Google Scholar
  86. Majors BS, Betenbaugh MJ, Pederson NE, Chiang GG (2009) Mcl-1 overexpression leads to higher viabilities and increased production of humanized monoclonal antibody in Chinese hamster ovary cells. Biotechnol Prog 25:1161–1168Google Scholar
  87. Mazur X, Fussenegger M, Renner WA, Bailey JE (1998) Higher productivity of growth-arrested Chinese hamster ovary cells expressing the cyclin-dependent kinase inhibitor p27. Biotechnol Prog 14:705–713Google Scholar
  88. Meents H, Enenkel B, Eppenberger HM, Werner RG, Fussenegger M (2002a) Impact of coexpression and coamplification of sICAM and antiapoptosis determinants bcl-2/bcl-x(L) on productivity, cell survival, and mitochondria number in CHO-DG44 grown in suspension and serum-free media. Biotechnol Bioeng 80:706–716Google Scholar
  89. Meents H, Enenkel B, Werner RG, Fussenegger M (2002b) p27Kip1-mediated controlled proliferation technology increases constitutive sICAM production in CHO-DUKX adapted for growth in suspension and serum-free media. Biotechnol Bioeng 79:619–627Google Scholar
  90. Meleady P, Henry M, Gammell P, Doolan P, Sinacore M, Melville M, Francullo L, Leonard M, Charlebois T, Clynes M (2008) Proteomic profiling of CHO cells with enhanced rhBMP-2 productivity following co-expression of PACEsol. Proteomics 8:2611–2624Google Scholar
  91. Mohan C, Park SH, Chung JY, Lee GM (2007) Effect of doxycycline-regulated protein disulfide isomerase expression on the specific productivity of recombinant CHO cells: thrombopoietin and antibody. Biotechnol Bioeng 98:611–615Google Scholar
  92. Mohan C, Kim YG, Koo J, Lee GM (2008) Assessment of cell engineering strategies for improved therapeutic protein production in CHO cells. Biotechnol J 3:624–630Google Scholar
  93. Müller D, Katinger H, Grillari J (2008) MicroRNAs as targets for engineering of CHO cell factories. Trends Biotechnol 26:359–365Google Scholar
  94. Nissom PM, Sanny A, Kok YJ, Hiang YT, Chuah SH, Shing TK, Lee YY, Wong KT, Hu WS, Sim MY, Philp R (2006) Transcriptome and proteome profiling to understanding the biology of high productivity CHO cells. Mol Biotechnol 34:125–140Google Scholar
  95. O'Callaghan PM, James DC (2008) Systems biotechnology of mammalian cell factories. Brief Funct Genomic Proteomic 7:95–110Google Scholar
  96. Ohya T, Hayashi T, Kiyama E, Nishii H, Miki H, Kobayashi K, Honda K, Omasa T, Ohtake H (2008) Improved production of recombinant human antithrombin III in Chinese hamster ovary cells by ATF4 overexpression. Biotechnol Bioeng 100:317–324Google Scholar
  97. Omasa T, Takami T, Ohya T, Kiyama E, Hayashi T, Nishii H, Miki H, Kobayashi K, Honda K, Ohtake H (2008) Overexpression of GADD34 enhances production of recombinant human antithrombin III in Chinese hamster ovary cells. J Biosci Bioeng 106:568–573Google Scholar
  98. Omasa T, Cao Y, Park JY, Takagi Y, Kimura S, Yano H, Honda K, Asakawa S, Shimizu N, Ohtake H (2009) Bacterial artificial chromosome library for genome-wide analysis of Chinese hamster ovary cells. Biotechnol Bioeng 104:986–994Google Scholar
  99. Park H, Kim IH, Kim IY, Kim KH, Kim HJ (2000) Expression of carbamoyl phosphate synthetase I and ornithine transcarbamoylase genes in Chinese hamster ovary dhfr-cells decreases accumulation of ammonium ion in culture media. J Biotechnol 81:129–140Google Scholar
  100. Pascoe DE, Arnott D, Papoutsakis ET, Miller WM, Andersen DC (2007) Proteome analysis of antibody-producing CHO cell lines with different metabolic profiles. Biotechnol Bioeng 98:391–410Google Scholar
  101. Peng RW, Fussenegger M (2009) Molecular engineering of exocytic vesicle traffic enhances the productivity of Chinese hamster ovary cells. Biotechnol Bioeng 102:1170–1181Google Scholar
  102. Peng RW, Abellan E, Fussenegger M (2011) Differential effect of exocytic SNAREs on the production of recombinant proteins in mammalian cells. Biotechnol Bioeng 108:611–620Google Scholar
  103. Renner WA, Lee KH, Hatzimanikatis V, Bailey JE, Eppenberger HM (1995) Recombinant cyclin E expression activates proliferation and obviates surface attachment of Chinese hamster ovary (CHO) cells in protein-free medium. Biotechnol Bioeng 47:476–482Google Scholar
  104. Ron D, Walter P (2007) Signal integration in the endoplasmic reticulum unfolded protein response. Nat Rev Mol Cell Biol 8:519–529Google Scholar
  105. Sauerwald TM, Betenbaugh MJ, Oyler GA (2002) Inhibiting apoptosis in mammalian cell culture using the caspase inhibitor XIAP and deletion mutants. Biotechnol Bioeng 77:704–716Google Scholar
  106. Sauerwald TM, Oyler GA, Betenbaugh MJ (2003) Study of caspase inhibitors for limiting death in mammalian cell culture. Biotechnol Bioeng 81:329–340Google Scholar
  107. Shen D, Kiehl TR, Khattak SF, Li ZJ, He A, Kayne PS, Patel V, Neuhaus IM, Sharfstein ST (2010) Transcriptomic responses to sodium chloride-induced osmotic stress: a study of industrial fed-batch CHO cell cultures. Biotechnol Prog 26:1104–1115Google Scholar
  108. Silva G, Poirot L, Galetto R, Smith J, Montoya G, Duchateau P, Pâques F (2011) Meganucleases and other tools for targeted genome engineering: perspectives and challenges for gene therapy. Curr Gene Ther 11:11–27Google Scholar
  109. Sung YH, Lee JS, Park SH, Koo J, Lee GM (2007) Influence of co-down-regulation of caspase-3 and caspase-7 by siRNAs on sodium butyrate-induced apoptotic cell death of Chinese hamster ovary cells producing thrombopoietin. Metab Eng 9:452–464Google Scholar
  110. Sunley K, Butler M (2010) Strategies for the enhancement of recombinant protein production from mammalian cells by growth arrest. Biotechnol Adv 28:385–394Google Scholar
  111. Tabuchi H, Sugiyama T, Tanaka S, Tainaka S (2010) Overexpression of taurine transporter in Chinese hamster ovary cells can enhance cell viability and product yield, while promoting glutamine consumption. Biotechnol Bioeng 107:998–1003Google Scholar
  112. Tey BT, Singh RP, Piredda L, Piacentini M, Al-Rubeai M (2000) Influence of bcl-2 on cell death during the cultivation of a Chinese hamster ovary cell line expressing a chimeric antibody. Biotechnol Bioeng 68:31–43Google Scholar
  113. Tigges M, Fussenegger M (2006) Xbp1-based engineering of secretory capacity enhances the productivity of Chinese hamster ovary cells. Metab Eng 8:264–272Google Scholar
  114. Timms JF, Cramer R (2008) Difference gel electrophoresis. Proteomics 8:4886–4897Google Scholar
  115. Toonen RF, Verhage M (2003) Vesicle trafficking: pleasure and pain from SM genes. Trends Cell Biol 13:177–186Google Scholar
  116. Van Dyk DD, Misztal DR, Wilkins MR, Mackintosh JA, Poljak A, Varnai JC, Teber E, Walsh BJ, Gray PP (2003) Identification of cellular changes associated with increased production of human growth hormone in a recombinant Chinese hamster ovary cell line. Proteomics 3:147–156Google Scholar
  117. Walsh G (2010) Biopharmaceutical benchmarks 2010. Nat Biotechnol 28:917–924Google Scholar
  118. Wlaschin KF, Hu WS (2007) Engineering cell metabolism for high-density cell culture via manipulation of sugar transport. J Biotechnol 131:168–176Google Scholar
  119. Wong DC, Wong KT, Lee YY, Morin PN, Heng CK, Yap MG (2006a) Transcriptional profiling of apoptotic pathways in batch and fed-batch CHO cell cultures. Biotechnol Bioeng 94:373–382Google Scholar
  120. Wong DC, Wong KT, Nissom PM, Heng CK, Yap MG (2006b) Targeting early apoptotic genes in batch and fed-batch CHO cell cultures. Biotechnol Bioeng 95:350–361Google Scholar
  121. Xu X, Nagarajan H, Lewis NE, Pan S, Cai Z, Liu X, Chen W, Xie M, Wang W, Hammond S, Andersen MR, Neff N, Passarelli B, Koh W, Fan HC, Wang J, Gui Y, Lee KH, Betenbaugh MJ, Quake SR, Famili I, Palsson BO, Wang J (2011) The genomic sequence of the Chinese hamster ovary (CHO)-K1 cell line. Nat Biotechnol 29:735–741. doi: 10.1038/nbt.1932 Google Scholar
  122. Yang M, Butler M (2000) Effects of ammonia on CHO cell growth, erythropoietin production, and glycosylation. Biotechnol Bioeng 68:370–380Google Scholar
  123. Yee JC, de Leon Gatti M, Philp RJ, Yap M, Hu WS (2008) Genomic and proteomic exploration of CHO and hybridoma cells under sodium butyrate treatment. Biotechnol Bioeng 99:1186–1204Google Scholar
  124. Yee JC, Gerdtzen ZP, Hu WS (2009) Comparative transcriptome analysis to unveil genes affecting recombinant protein productivity in mammalian cells. Biotechnol Bioeng 102:246–263Google Scholar
  125. Yoon SK, Hwang SO, Lee GM (2004) Enhancing effect of low culture temperature on specific antibody productivity of recombinant Chinese hamster ovary cells: clonal variation. Biotechnol Prog 20:1683–1688Google Scholar
  126. Yun CY, Liu S, Lim SF, Wang T, Chung BY, Jiat Teo J, Chuan KH, Soon AS, Goh KS, Song Z (2007) Specific inhibition of caspase-8 and −9 in CHO cells enhances cell viability in batch and fed-batch cultures. Metab Eng 9:406–418Google Scholar
  127. Zahn-Zabal M, Kobr M, Girod PA, Imhof M, Chatellard P, de Jesus M, Wurm F, Mermod N (2001) Development of stable cell lines for production or regulated expression using matrix attachment regions. J Biotechnol 87:29–42Google Scholar
  128. Zhang F, Sun X, Yi X, Zhang Y (2006) Metabolic characteristics of recombinant Chinese hamster ovary cells expressing glutamine synthetase in presence and absence of glutamine. Cytotechnology 51:21–28Google Scholar
  129. Zhou M, Crawford Y, Ng D, Tung J, Pynn AF, Meier A, Yuk IH, Vijayasankaran N, Leach K, Joly J, Snedecor B, Shen A (2011) Decreasing lactate level and increasing antibody production in Chinese Hamster Ovary cells (CHO) by reducing the expression of lactate dehydrogenase and pyruvate dehydrogenase kinases. J Biotechnol 153:27–34Google Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  1. 1.Department of Biological SciencesGraduate School of Nanoscience & Technology (WCU), KAISTDaejeonKorea
  2. 2.Biotechnology Process Engineering Center, KRIBBDaejeonKorea

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