Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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High-Level Expression of Recombinant Human Interleukin-2 in Chinese Hamster Ovary Cells Using the Expression System Containing Transcription Terminator

  • Published : 2004.08.01
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Abstract

Many biological properties and the clinical potential of human interleukin-2 (hIL-2) draw much attention to its high-level expression in mammalian cells. Recombinant human IL-2 (rhIL-2) was expressed in Chinese hamster ovary (CHO) cells, using the recently developed expression system which confers position-independent expression. Stable CHO cell lines carrying several hundred amplified copies of the rhIL-2 gene were easily obtained and rhIL-2 was expressed at high levels after selection with increasing concentrations of methotrexate. Interestingly, the insertion of the transcription terminator of the human gastrin gene into the downstream region of the gene for rhIL-2 considerably increased rhIL-2 expression. Using the expression system with the transcription terminator, it was possible to get a CHO cell line expressing the rhIL-2 at a very high level, about $11.4\mug/10^6$ cell/day, which is about 6 times higher than that previously reported. The biological activity of the rhIL-2 protein purified from the cell line was also confirmed by the cell proliferation assay.

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References

  1. Proc. Natl. Acad. USA v.83 RNA polymerase Ⅱ transcription terminates at a specific DNA sequence in a HeLa cell-free reaction Baek, K.;K. Sato;R. Ito;K. Agarwal https://doi.org/10.1073/pnas.83.20.7623
  2. Proc. Natl. Acad. Sci. USA v.83 Identification of a common nucleotide sequence in the 3'-untranslated region of mRNA molecules specifying inflammatory mediators Caput, D.;B. Beutler;K. Hartog;R. Thayer;S. Brown-Shimer;A. Cerami https://doi.org/10.1073/pnas.83.6.1670
  3. J. Exp. Med. v.155 Augmentation of the anti-tumor therapeutic efficacy of long-term cultured T lymphocytes by in vivo administration of purified interleukin 2 Cheever, M. A.;P. D. Fefer;S. Gills https://doi.org/10.1084/jem.155.4.968
  4. Cell v.107 AU binding proteins recruit the exosome to degrade ARE-containing mNAs Chen, C.;R. Gherzi;S. Ong;E. Chan;R. Raijmakers https://doi.org/10.1016/S0092-8674(01)00578-5
  5. J. Microbiol. Biotechnol. v.12 Screening of high-porductivity cell lines and investigation of their physiology in Chinese hamster ovary (CHO) cell cultures for transforming growth factor-${\beta}$ production Chun, G. T.;J. B. Lee;S. U. Nam;S. W. Lee;Y. H. Jeong;E. Y. Choi;I. H. Kim;Y. S. Jeong;P. H. Kim
  6. J. Biol. Chem. v.263 Deletion analysis of the Chinese hamster dihydrofolate reductase ene promoter Ciudad, C. K.;G. Urlaub;L. A. Chasin
  7. Eur. J. Biochem. v.153 Structures of the major carbohydrates of natural human interleukin-2 Conradt, H. S.;R. Geyer;J. Hoppe;L. Grotjahn;A. Plessing;H. Mohr https://doi.org/10.1111/j.1432-1033.1985.tb09295.x
  8. Nucleic Acids Res. v.11 Molecular cloning of human interleukin 2 cDNA and its expression in E. coli Devos, R.;G. Plaetinck;H. Cheroutre;G. Simons;W. Degrave;J. Tavernier;E. Remaut;W. Fiers https://doi.org/10.1093/nar/11.13.4307
  9. J. Immunol. v.130 Interleukin 2 dependence of human natural killer (NK) cell activity Domzing, W.;B. M. Stadler;R. B. Herberman
  10. J. Immunol. v.126 Regulation of the production of immune interferon and cytotoxic T lymphocytes by interleukin 2 Farrar, W. L.;H. M. Johnson;J. J. Farrar
  11. FEBS Lett. v.226 Characterization of recombinant glycosylated human interleukin 2 produced by a recombinant plasmid transformed CHO cell line Ferrara, P.;E. Pecceu,E. Marchese;N. Vita;W. Roskam;J. Lupker https://doi.org/10.1016/0014-5793(87)80548-3
  12. J. Biol. Chem. v.276 Interleukin-2 carbohydrate recognition modulates CTLL-2 cell proliferation Fukushima, K.;K. Yamashita https://doi.org/10.1074/jbc.M008781200
  13. J. Exp. Med. v.155 Lymphokine-activated killer cell phenomenon. Lysis of naturalkill-resistant fresh solid tumor cells by interleukin 2-activated autologous human peripheral blood lymphocytes Grimm, E. A.;A. Mazumber,H. Z. Zhang;S. A. Rogenberg https://doi.org/10.1084/jem.155.6.1823
  14. Biotechnol. Prog. v.19 Improved mammalian expression systems by manipulating trancriptional temination regions Kim, D.;J. D. Kim;K. Baek;Y. Yoon;J. Yoon https://doi.org/10.1021/bp0341186
  15. J. Biotechnol. v.107 Improved recombinant gene expression in CHO cells using matrix attachment regions Kim, J. M.;J. S. Kim;D. H. Park;H. S. Kang;J. Yoon;K. Baek;Y. Yoon https://doi.org/10.1016/j.jbiotec.2003.09.015
  16. J. Microbiol. Biotechnol. v.13 Characterization of humanized antibody produced by apoptosis-resistant CHO cells under sodium bytyrate-induced condition Kim, N. S.;K. H. Chang;B. S. Chung;S. H. Kim;J. H. Kim;G. M. Lee
  17. Nucleic Acids Res. v.4 Compilation and anlysis of sequences upstream from the translational start site in eukaryotic mRNAs Kozak, M.
  18. Biochem. J. v.229 Characterization of human interleukin 2 derived from Escherichia coli Liang, S. M.;B. Allet;K. Rose;M. Hirschi;C. M. Liang;D. R. Thatcher https://doi.org/10.1042/bj2290429
  19. J. Chromatogr v.504 Separation by cation-exchange high-performance liquid chromatography of three forms of Chinese hamster ovary cell-derived recombinant human interleukin-2 Marchese, E.;N. Vita;T. Maureaud;P. Ferrara https://doi.org/10.1016/S0021-9673(01)89538-8
  20. Science v.193 Selective in vitro growth of T lymphocytes from normal human bone marrows Morgan, D. A.;F. W. Ruscetti;R. Gallo https://doi.org/10.1126/science.181845
  21. Immunol. Today v.17 The expanding universe of T-cell subsets: Th1, Th2 and more Mosmann, T. R.;S. Sad https://doi.org/10.1016/0167-5699(96)80606-2
  22. Science v.225 Adoptive immunotherapy of established pulmonary metastases with LAK cells and recombinant interleukin-2 Mule, J. J.;S. Shu;S. L. Schwarz;S. A. Rosenberg https://doi.org/10.1126/science.6332379
  23. Biochem. Biophys. Res. Commun. v.128 Comparison of the biological properties of purified natural and recombinant human interleukin-2 Naruo, K.;S. Hinuma,K. Kato;M. Koyama;H. Tada;O. Shiho;K. Tsukamoto https://doi.org/10.1016/0006-291X(85)91672-9
  24. J. Biochem. v.102 Expession of amplified cDNA and genomic sequnces encoding human interleukin 2 in Chinese hamster ovary cells Onomichi, K.;Y. Eto;H. Shibai https://doi.org/10.1093/oxfordjournals.jbchem.a122023
  25. Immunobiology v.174 Relative cloning efficiencies and long-term propagation capacity for T cell clones of highly purified natual interleukin 2 compared to recombinant interleukin 2 in man Pawelec, G.;U. Schwulera;M. Blaurock;F. W. Busch;A. Rehbein;I. Balko;P. Wernet https://doi.org/10.1016/S0171-2985(87)80085-2
  26. Nature v.322 Transcription interference and termination between duplicated ${\alpha}$-globin gene constructs suggests a novel mechanism for gene regulation Proudfoot, N. J. https://doi.org/10.1038/322562a0
  27. Proc. Natl. Acad. Sci. USA v.81 Amino acid squence and posttranslational modification of human interleukin 2 Robb, R. J.;R. M. Kutny;M. Panico;H. R. Morris;V. Chowdhry https://doi.org/10.1073/pnas.81.20.6486
  28. Science v.223 Biological activity of recombinant human interleukin-2 produced in Escherichia coli Rosenberg, S. A.;E. A. Grimm;M. McGrogan;M. Doyle;E. Kawasaik;K. Koths;D. F. Mark https://doi.org/10.1126/science.6367046
  29. Molecular Cloning: A Laboratory Manual Sambrook, J.;E. F. Fritsch;T. Maniaits
  30. Nature v.302 Structure and expression of a cloned cDNA for human interleukin-2 Taniguchi, T.;H. Matsui;T. Fujita;C. Takaoka;N. Kashima;R. Yoshimoto;J. Hamuro https://doi.org/10.1038/302305a0
  31. Science v.224 Site-specific mutagenesis of the human interleukin-2 gene: Structure-function analysis of the cysteine residues Wang, A.;S. D. Lu;D. F. Mark https://doi.org/10.1126/science.6427925