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Applied Biochemistry and Biotechnology

, Volume 163, Issue 1, pp 1–13 | Cite as

Behavior of Wild-type and Transfected S2 Cells Cultured in Two Different Media

  • Fabiana R. X. Batista
  • Kátia N. Greco
  • Renato M. Astray
  • Soraia A. C. Jorge
  • Elisabeth F. P. Augusto
  • Carlos A. Pereira
  • Ronaldo Z. Mendonça
  • Ângela M. MoraesEmail author
Article

Abstract

An animal protein-free medium composed of IPL-41 containing 6 g L−1 yeastolate ultrafiltrate, 10 g L−1 glucose, 2 g L−1 lactose, 5 g L−1 glutamine, 1% lipid emulsion, and 0.1% Pluronic F-68 was used for producing recombinant proteins in batch mode employing two cell lines, S2AcRVGP2k expressing the G glycoprotein from rabies virus (RVGP) and S2AcHBsAgHy-9C expressing the surface antigen of hepatitis B virus (HBsAg), both obtained from Drosophila melanogaster S2 cells. Growth of wild-type S2 cells was also evaluated in the same medium. Cell behavior in the tested medium was compared to that verified in Sf900 II®. The results show that in shake flasks, S2AcRVGP2k and S2AcHBsAgHy-9C cells reached around 2 × 107 cells mL−1 in both media. In supplemented IPL-41 and Sf900 II® media, S2AcRVGP2k cells produced 367 ng RVGP mL−1 and 638 ng RVGP mL−1, respectively, while S2AcHBsAgHy-9C cells correspondently produced 573 ng HBsAg mL−1 and 322 ng HBsAg mL−1 in the mentioned media. In stirred tanks, S2AcRVGP2k cells reached 3 × 107 cells mL−1 and produced up to 758 ng RVGP mL−1. In general, glucose was consumed by cells, while lactate and ammonia were produced.

Keywords

S2 cells Metabolism RVGP HBsAg Animal protein-free medium 

Notes

Acknowledgments

The authors acknowledge Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, Brasília, Brazil), Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP, grant no. 02/09482-3, São Paulo, Brazil), Instituto de Pesquisas Tecnológicas do Estado de São Paulo (IPT, São Paulo, Brazil) and Instituto Butantan (São Paulo, Brazil) for the financial support. The authors gratefully acknowledge M Aguiar and R Andrade (IPT, São Paulo, Brazil) for the technical assistance. AM Moraes and CA Pereira are recipients of CNPq fellowships.

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Fabiana R. X. Batista
    • 1
  • Kátia N. Greco
    • 2
  • Renato M. Astray
    • 2
  • Soraia A. C. Jorge
    • 2
  • Elisabeth F. P. Augusto
    • 3
  • Carlos A. Pereira
    • 2
  • Ronaldo Z. Mendonça
    • 2
  • Ângela M. Moraes
    • 1
    Email author
  1. 1.Departamento de Processos Biotecnológicos, Faculdade de Engenharia QuímicaUniversidade Estadual de CampinasCampinasBrazil
  2. 2.Laboratório de Imunologia ViralInstituto ButantanSão Paulo-SPBrazil
  3. 3.Laboratório de Biotecnologia IndustrialInstituto de Pesquisas Tecnológicas do Estado de São PauloSão Paulo-SPBrazil

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