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Optimization of growth methods and recombinant protein-production in bti-tn-5b1-4 insect cells using the baculovirus expression system

Academic Article
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Overview

authors

  • Wickham, T. J.
  • Nemerow, Glen

publication date

  • January 1993

journal

  • Biotechnology Progress  Journal

abstract

  • A novel insect cell line from Trichoplusia ni, BTI-Tn 5B1-4 (Tn 5), was compared to Spodoptera frugiperda, Sf 9, cells for production of two recombinant secreted proteins: truncated Epstein-Barr viral attachment protein (EBV gp105) and truncated, soluble tissue factor (sTF). Under optimum conditions for both cell lines, Tn 5 cells produced 28-fold more secreted sTF than Sf 9 cells, respectively, on a per cell basis. The total production of gp105 was similar for the two cell lines. However, Tn5 cells secreted gp105 much more efficiently, resulting in 5-fold higher levels in the extracellular medium. Despite these increases, Tn 5 cells are attachment-dependent, and protein production is sensitive to the cell density (cells/cm2), unlike the Sf9 cell line which can be easily grown and scaled up in cell suspension cultures without significantly affecting its per cell production. Thus, protein production from Tn 5 cells above 0.1 L scales was optimized with respect to cell density using standard techniques for the growth of attachment-dependent cells. Roller bottles precoated with DEAE-based microcarriers and suspension cultures employing collagen-coated microcarriers were found to be effective ways of culturing Tn 5 cells. Predetermined optimal cell densities were used to produce EBV gp105 in microcarrier-coated roller bottles or in suspension cultures using collagen-coated microcarriers at concentrations close to those observed in tissue culture flasks.

subject areas

  • Antigens, Viral
  • Baculoviridae
  • Cell Division
  • Cell Line
  • Electrophoresis, Polyacrylamide Gel
  • Genetic Vectors
  • Herpesvirus 4, Human
  • Recombinant Proteins
  • Thromboplastin
  • Viral Envelope Proteins
  • Viral Matrix Proteins
  • beta-Galactosidase
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Identity

International Standard Serial Number (ISSN)

  • 8756-7938

Digital Object Identifier (DOI)

  • 10.1021/bp00019a004

PubMed ID

  • 7764044
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Additional Document Info

start page

  • 25

end page

  • 30

volume

  • 9

issue

  • 1

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