Skip to main content
SpringerLink
Log in

Effect of culture temperature on a recombinant CHO cell line producing a C-terminal α-amidating enzyme

  • Published:
Cytotechnology Aims and scope Submit manuscript

Abstract

In order to seek an efficient method for producing a recombinant protein by using animal cell culture, we investigated various effects of the culture temperature on a recombinant CHO cell line (3µ-1S), producing a C-terminal α-amidating enzyme (799BglIIα-AE) originating from Xenopus laevis. The results revealed that a low culture temperature (below 37 °C) led to the following phenomena: [1] inhibited cell growth, [2] enhanced cellular productivity of the recombinant protein, [3] maintained high cell viability, [4] suppressed medium consumption, and [5] suppressed release of impurities from the cells. These findings indicate that a quite simple method, the culture at low temperature, will contribute to the total improvement of the industrial process for the production of the recombinant protein, 799BglIIα-AE.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Bloemkolk J-W, Gray MR, Merchant F and Mosmann TR (1992) Effect of temperature on hybridoma cell cycle and MAb production. Biotechnol Bioeng 40: 427-431.

    Article  CAS  Google Scholar 

  • Borth N, Heider R, Assadian A and Katinger H (1992) Growth and production kinetics of human x mouse and mouse hybridoma cells at reduced temperature and serum content. J Biotechnol 25: 319-331.

    Article  CAS  Google Scholar 

  • Borys MC, Linzer DIH and Papoutsakis ET (1993) Culture pH affects expression rates and glycosylation of recombinant mouse placental lactogen proteins by Chinese hamster ovary (CHO) cells. Bio/Technol 11: 720-724.

    Article  CAS  Google Scholar 

  • Borys MC, Linzer DIH and Papoutsakis ET (1994) Ammonia affects the glycosylation patterns of recombinant mouse placental lactogenI by Chinese hamster ovary cells in a pH-dependent manner. Biotechnol Bioeng 43: 505-514.

    Article  CAS  Google Scholar 

  • Bradbury AF, Finnie MDA and Smyth GG (1982) Mechanism of C-terminal amide formation by pituitary enzymes. Nature 305: 686-688.

    Article  Google Scholar 

  • Eiper BA, Mains RE and Glembotski CC (1983) Identification in pituitary tissue of a peptide α-amidation activity that acts on glycine-extended peptides and requires molecular oxygen, copper, and ascorbic acid. Proc Natl Acad Sci USA 80: 5144-5148.

    Article  Google Scholar 

  • AE) in recombinant CHO cells}. In: Kaminogawa, S et al. (eds.) Animal Cell Technology: Basic and Applied Aspects, Vol. 5(pp. 493-499) Kluwer Academic Publishers, The Netherlands.

    Google Scholar 

  • Giard DJ and Fleischaker RJ (1980) Examination of parameters affecting human interferon production with microcarrier-grown fibroblast cells. Antimicrob Agents Chemother 18: 130-136.

    CAS  Google Scholar 

  • Giard DJ, Fleischaker RJ and Fabricant M (1982) Effect of temperature on the production of human fibroblast interferon (41411). Proc Soc Exp Biol Med 170: 155-159.

    CAS  Google Scholar 

  • Goldblum S, Bae YK, Hink WF and Chalmers J (1990) Protective effect of methylcellulose and other polymers on insect cells subjected to laminar shear stress. Biotechnol Prog 6: 383-390.

    Article  CAS  Google Scholar 

  • Goochee CF and Monica T (1990) Environmental effects on protein glycosylation. Bio/Technol 8: 421-427.

    Article  CAS  Google Scholar 

  • Hahn TJ and Goochee CF (1992) Growth-associated glycosylation of transferrin secreted by HepG2 cells. J Biol Chem 267: 23982-23987.

    CAS  Google Scholar 

  • Hayter PM, Curling EMA, Baines AJ, Jenkins N, Salmon I, Strange PG, Tong JM and Bull AT (1992) Glucose-limited chemostat culture of Chinese hamster ovary cells producing recombinant human interferon-γ. Biotechnol Bioeng 39: 327-335.

    Article  CAS  Google Scholar 

  • Hosoi S, Murosumi K, Sasaki K, Satoh M, Miyaji H, Hasegawa M, Itoh S, Tamaoki T and Sato S (1991) Optimization of cell culture conditions for GCSF (granulocyte-colony stimulating factor) production by genetically engineered Namalwa KJM-1 cells. Cytotechnology 7: 25-32.

    Article  CAS  Google Scholar 

  • Kato I, Yonekura H, Tajima M, Yanagi M, Yamamoto H and Okamoto H (1990) Two enzymes concerned in peptide hormone-amidation are synthesized from a single mRNA. Biochem Biophys Res Comm 172: 197-203.

    Article  CAS  Google Scholar 

  • Kojima Y and Yoshida F (1974) Enhanced production of interferon by temperature shift-down from 37 °C to 25 °C in rabbit cell cultures stimulated with Newcastle disease virus. Japan. J Microbiol 18: 217-222.

    CAS  Google Scholar 

  • Lin AA, Kimura R and Miller WM (1993) Production of tPA in recombinant CHO cells under oxygen-limited conditions. Biotechnol Bioeng 42: 339-350.

    Article  CAS  Google Scholar 

  • McQueen A, Meilhoc E and Bailey JE (1987) Flow effects on the viability and lysis of suspended mammalian cells. Biotechnol Lett 9: 831-836.

    Article  CAS  Google Scholar 

  • Merten OW (1988) Batch production and growth kinetics of hybridomas. Cytotechnology 1: 113-121.

    Article  Google Scholar 

  • Mizuno K, Ohsuye K, Wada Y, Fuchimura K, Tanaka S and Matsuo H (1987) Cloning and sequence of cDNA encoding a peptide C-terminal α-amidating enzyme from Xenopus laevis. Biochem. Biophys. Res. Comm. 148: 546-552.

    Article  CAS  Google Scholar 

  • Mizuno K, Sakata J, Kojima M, Kangawa K and Matsuo H (1986) Peptide C-terminal α-amidating enzyme purified to homogeneity from Xenopus laevisskin. Biochem Biophys Res Comm 137: 984-991.

    Article  CAS  Google Scholar 

  • Mochizuki K, Sato S, Kato M and Hashizume S (1993) Enhanced production of human monoclonal antibodies by the use of fructose in serum-free hybridoma culture media. Cytotechnology 13: 161-173.

    Article  CAS  Google Scholar 

  • Ohsuye K, Kitano K, Wada Y, Fuchimura K, Tanaka S, Mizuno K and Matsuo H (1988) Cloning of cDNA encoding a new peptide C-terminal αamidating enzyme having a putative membrane-spanning domain from Xenopus laevisskin. Biochem Biophys Res Comm 150: 1275-1281.

    Article  CAS  Google Scholar 

  • Ozturk SS and Palsson BO (1991) Growth, metabolic, and antibody production kinetics of hybridoma cell culture: 2. Effects of serum concentration, dissolved oxygen concentration, and medium pH in a batch reactor. Biotechnol Prog 7: 481-494.

    Article  CAS  Google Scholar 

  • Patel TP, Parekh RB, Moellering BJ and Prior CP (1992) Different culture methods lead to differences in glycosylation of a murine IgG monoclonal antibody. Biochem J 285: 839-845.

    CAS  Google Scholar 

  • Petersen JF, McIntire LV and Papoutsakis ET (1988) Shear sensitivity of cultured hybridoma cells (CRL8018) depends on mode of growth, culture age and metabolite concentration. J Biotechnol 7: 229-246.

    Article  CAS  Google Scholar 

  • Reuveny S, Velez D, Macmillan JD and Miller L (1986) Factors affecting cell growth and monoclonal antibody production in stirred reactors. J Immunol Methods 86: 53-59.

    Article  CAS  Google Scholar 

  • Sureshkumar GK and Mutharasan R (1991) The influence of temperature on a mouse-mouse hybridoma growth and monoclonal antibody production. Biotechnol Bioeng 37: 292-295.

    Article  CAS  Google Scholar 

  • Tachibana H, Jiyoun K, Taniguchi K, Ushio Y, Teruya K, Osada K, Inoue Y, Shirahata S and Murakami H (1996) Modified antigen-binding of human antibodies with glycosylation variations of the light chains produced in sugar-limited human hybridoma cultures. In Vitro Cell Dev Biol 32: 178-183.

    CAS  Google Scholar 

  • Takagi M and Ueda K(1994) Comparison of the optimal culture conditions for cell growth and tissue plasminogen activator production by human embryo lung cells on microcarriers. Appl Microbiol Biotechnol 41: 565-570.

    Article  CAS  Google Scholar 

  • Vilček J and Havell EA (1973) Stabilization of interferon messenger RNA activity by treatment of cells with metabolic inhibitors and lowering of the incubation temperature. Proc Natl Acad Sci USA 70: 3909-3913.

    Article  Google Scholar 

  • Weidemann R, Ludwig A and Kretzmer G (1994) Low temperature cultivation - A step towards process optimization. Cytotechnology 15: 111-116.

    Article  CAS  Google Scholar 

  • Wu J, Daugulis AJ, Faulkner P and Goosen MFA (1992) Correlation of LDH activity with loss of insect cell viability: An assessment of the LDH assay. Biotechnol Tech 6: 335-340.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Suntory Institute for Medicinal Research and Development, 2716-1 Kurakake, Akaiwa, Chiyoda-machi, Ohra-gun, Gunma, 370-05, Japan

    Kazuaki Furukawa & Kazuhiro Ohsuye

Authors
  1. Kazuaki Furukawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kazuhiro Ohsuye
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Furukawa, K., Ohsuye, K. Effect of culture temperature on a recombinant CHO cell line producing a C-terminal α-amidating enzyme. Cytotechnology 26, 153–164 (1998). https://doi.org/10.1023/A:1007934216507

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1007934216507

Search

Navigation