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Comparative characterization of growth and recombinant protein production among three insect cell lines with four kinds of serum free media

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Abstract

Three insect cell lines, Sf9, Sf21 and Tn5B1-4, and four different kinds of serum free media (SFM), Sf 900 II, EX-CELL 420, EX-CELL 405 and Express Five, were used to compare the nutrient consumption, byproduct formation, production of recombinant protein and protease activity in suspension cultures. The Sf 900 II SFM was appropriate for the cell growth and protein production of the Sf9 and Sf21 cell lines. When the Tn5B1-4 cell line was grown in the Express Five SFM, the specific growth rate was 1.6 fold higher than those of either the Sf9 or Sf21 cell lines. The glucose and glutamine consumption rates per cells, were 4 and 2.3. times higher than those of the Sf9 cell line, respectively. The overall yield coefficients of the lactate and ammonium ion were 2.8 and 1.5 times higher compared to those of the Sf9 cell line, respectively. The maximum specific β-galactosidase production rate was 4.5. fold that of the Sf9 cell line, a 3 times higher protease activity per cell.

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References

  1. [1]

    Luckow, V. A. (1995) Protein production and processing from baculovirus expression vectors. pp. 51–90. In: M. L. Shuler, H. A. Wood, R. R. Granados, and D. A. Hammer, (eds.).Baculovirus Expression System and Biopesticides, Wiley-Liss, New York, USA.

  2. [2]

    Palomares, L. A. and O. T. Ramirez (1997) Insect cell culture: recent advances, bioengineering challenges and implications in protein production. pp. 25–52. In: E. Galindo and O. T. Ramirez (eds.).Advances in Bioprocess Engineering II. Kluwer Academic Publishers, Dordrecht, The Netherlands.

  3. [3]

    Agathos, S. N., Y. H. Jeong, and K. Venkat (1990) Growth kinetics of free and immobilized insect cell cultures.Ann. N. Y. Acad. Sci. 589: 372–398.

  4. [4]

    Maranga, L., P. E. Cruz, J. G. Aunins, and M. J. T. Carrondo (2002) Production of core and virus-like particles with baculovirus infected insect cells.Adv. Biochem. Eng. Biotechnol. 74: 183–206.

  5. [5]

    Donaldson, M. S. and M. L. Shuler (1998) Effects of long-term passing of BTI-Tn5B1-4 insect cells on growth and recombinant protein production.Biotechnol. Prog. 14: 543–547.

  6. [6]

    Donaldson, M. S. and M. L. Shuler (1998) Low-cost serum-free medium for the BTITn5B1-4 insect cell line.Biotechnol. Prog. 14: 573–579.

  7. [7]

    Miller, J. H. (1972) Assay of β-galactosidase. In:Experiments in Molecular Genetics. pp. 352–355. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, USA.

  8. [8]

    Reed, L. and H. Muench (1938) A simple method for estimating fifty percent endpoints.Am. J. Hyg. 27: 494–497.

  9. [9]

    Slack, J. M., J. Kuzio, and P. Faulkner (1995) Characterization of v-cath, a cathepsin L-like protease expressed by the baculovirusAutographa californica multiple nuclear polyhedrosis virus.J. Gen. Virol. 76: 1091–1098.

  10. [10]

    Ohkawa T., K. Majima, and S. Maeda (1994) A cystein protease encoded by the baculovirusBombyx mori nuclear polyhedrosis virus.J. Viol., 68: 6619–6625

  11. [11]

    Gotoh, T., Y. Miyazaki, K. I. Kikuchi, and W. E. Bentley (2001) Investigation of sequential behavior of carboxyl protease and cystein protease activities in virus-infected Sf-9 insect cell culture by inhibition assay.Appl. Microbiol. Biotechnol. 56: 742–749

  12. [12]

    Kato, T., E. Y. Park, T. Murata, and T. Usui (2003) Improvement of GFPuv-ß3GnT2 fusion protein production by suppressing protease in baculovirus expression system.J. Biosci. Bioeng. Submitted in 2003.

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Correspondence to Enoch Y. Park.

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Kwon, M.S., Dojima, T. & Park, E.Y. Comparative characterization of growth and recombinant protein production among three insect cell lines with four kinds of serum free media. Biotechnol Bioproc E 8, 142–146 (2003). https://doi.org/10.1007/BF02940270

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Keywords

  • baculovirus expression vector system
  • serum free medium
  • insect cell
  • β-galactosidase
  • protease