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Semi-stable production of bovine IL-4 and GM-CSF in the mammalian episomal expression system

Federico Carlos Blanco

Federico Carlos Blanco

  • Institute of Agrobiotechnology and Molecular Biology, National Institute of Agricultural Technology–National Scientific and Technical Research CouncilBuenos Aires, Argentina
  • Institute of Biotechnology, Centre for Veterinary Scientific and Agronomics Research, National Institute of Agricultural TechnologyBuenos Aires, Argentina
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Blanco, Federico Carlos
, 
Cristina Lourdes Vazquez

Cristina Lourdes Vazquez

  • Institute of Agrobiotechnology and Molecular Biology, National Institute of Agricultural Technology–National Scientific and Technical Research CouncilBuenos Aires, Argentina
  • Institute of Biotechnology, Centre for Veterinary Scientific and Agronomics Research, National Institute of Agricultural TechnologyBuenos Aires, Argentina
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Vazquez, Cristina Lourdes
, 
Julia Sabio y García

Julia Sabio y García

  • Institute of Agrobiotechnology and Molecular Biology, National Institute of Agricultural Technology–National Scientific and Technical Research CouncilBuenos Aires, Argentina
  • Institute of Biotechnology, Centre for Veterinary Scientific and Agronomics Research, National Institute of Agricultural TechnologyBuenos Aires, Argentina
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García, Julia Sabio y
, 
Rosana Valeria Rocha

Rosana Valeria Rocha

  • Institute of Agrobiotechnology and Molecular Biology, National Institute of Agricultural Technology–National Scientific and Technical Research CouncilBuenos Aires, Argentina
  • Institute of Biotechnology, Centre for Veterinary Scientific and Agronomics Research, National Institute of Agricultural TechnologyBuenos Aires, Argentina
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Rocha, Rosana Valeria
, 
María José Gravisaco

María José Gravisaco

  • Institute of Agrobiotechnology and Molecular Biology, National Institute of Agricultural Technology–National Scientific and Technical Research CouncilBuenos Aires, Argentina
  • Institute of Biotechnology, Centre for Veterinary Scientific and Agronomics Research, National Institute of Agricultural TechnologyBuenos Aires, Argentina
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Gravisaco, María José
, 
Marina Andrea Forrellad

Marina Andrea Forrellad

  • Institute of Agrobiotechnology and Molecular Biology, National Institute of Agricultural Technology–National Scientific and Technical Research CouncilBuenos Aires, Argentina
  • Institute of Biotechnology, Centre for Veterinary Scientific and Agronomics Research, National Institute of Agricultural TechnologyBuenos Aires, Argentina
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Forrellad, Marina Andrea
, 
Giovanni Magistrelli

Giovanni Magistrelli

Light Chain Bioscience – Novimmune SAGeneva, Switzerland
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Magistrelli, Giovanni
 and 
Fabiana Bigi

Fabiana Bigi

  • Institute of Agrobiotechnology and Molecular Biology, National Institute of Agricultural Technology–National Scientific and Technical Research CouncilBuenos Aires, Argentina
  • Institute of Biotechnology, Centre for Veterinary Scientific and Agronomics Research, National Institute of Agricultural TechnologyBuenos Aires, Argentina
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Bigi, Fabiana
  
Aug 19, 2021
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Published Online: Aug 19, 2021
Page range: 315 - 321
Received: Jan 22, 2021
Accepted: Jul 26, 2021
DOI: https://doi.org/10.2478/jvetres-2021-0043
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© 2021 F.C. Blanco et al. published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Fig. 1

PEAKrapid CRL-2828 cells transfected with plasmids expressing bovine IL-4 and bovine GM-CSF together with plasmids expressing GFP
PEAKrapid CRL-2828 cells transfected with plasmids expressing bovine IL-4 and bovine GM-CSF together with plasmids expressing GFP

Fig. 2

Expression of rIL-4 and rGM-CSF in culture supernatants of transfected cells. Western blot analysis of total secreted proteins from transfected cells. Total proteins were precipitated with 10 % TCA from culture supernatant of non-transfected cells (1) of cells transfected with plasmid expressing IL-4 (2 and 4) or GM-CSF (3and 5). Molecular weight marker was loaded on lane 6
Expression of rIL-4 and rGM-CSF in culture supernatants of transfected cells. Western blot analysis of total secreted proteins from transfected cells. Total proteins were precipitated with 10 % TCA from culture supernatant of non-transfected cells (1) of cells transfected with plasmid expressing IL-4 (2 and 4) or GM-CSF (3and 5). Molecular weight marker was loaded on lane 6

Fig. 3

Functional activity of culture supernatant of IL-4 and GM-CSF transfected cells. Bovine monocytes were incubated with 10% v/v cell culture supernatant containing rIL-4 and/or rGM-CSF. As a control, monocytes were incubated with 10% v/v cell culture supernatant of non-transfected cells. Cell morphologies were registered with a confocal microscope
Functional activity of culture supernatant of IL-4 and GM-CSF transfected cells. Bovine monocytes were incubated with 10% v/v cell culture supernatant containing rIL-4 and/or rGM-CSF. As a control, monocytes were incubated with 10% v/v cell culture supernatant of non-transfected cells. Cell morphologies were registered with a confocal microscope
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English
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Journal Subjects:
Life Sciences, Molecular Biology, Microbiology and Virology, Life Sciences, other, Medicine, Veterinary Medicine
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