Replacing factor-dependency with that for lysozyme: affordable culture of IL-6-dependent hybridoma by transfecting artificial cell surface receptor

Biotechnol Bioeng. 2001 Sep 5;74(5):416-23. doi: 10.1002/bit.1132.

Abstract

Cytokines and growth factors are indispensable for the propagation and maintenance of factor-dependent mammalian cells. However, cytokines are often so expensive that the use of factor-dependent cells for industrial applications such as protein production is often not practical. Based on our previous design of a binary hen egg lysozyme (HEL)-specific receptor composed of portions of the anti-HEL antibody and the erythropoietin receptor, a new pair of chimeric receptors having the intracellular domain of gp130 were made and transfected to an interleukin-6 (IL-6)-dependent hybridoma, 7TD1. The clone expressing the two new receptors showed clear HEL dose-dependent cell growth and monoclonal antibody production in both serum-based and serum-free media without IL-6. These results establish the feasibility of applying receptor design to tailor cells for the inexpensive induction of cell growth for the purpose of producing therapeutic products.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / metabolism
  • Cell Division / physiology
  • Cell Line / cytology
  • Cell Line / metabolism
  • Culture Media, Serum-Free / metabolism
  • DNA-Binding Proteins / metabolism*
  • Efficiency
  • Hybridomas / cytology
  • Hybridomas / metabolism*
  • Interleukin-6 / metabolism*
  • Mice
  • Muramidase / metabolism*
  • Phosphorylation
  • Receptors, Cell Surface / genetics
  • Receptors, Cell Surface / metabolism
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism*
  • STAT3 Transcription Factor
  • Trans-Activators / metabolism*
  • Transfection

Substances

  • Antibodies, Monoclonal
  • Culture Media, Serum-Free
  • DNA-Binding Proteins
  • Interleukin-6
  • Receptors, Cell Surface
  • Recombinant Fusion Proteins
  • STAT3 Transcription Factor
  • Stat3 protein, mouse
  • Trans-Activators
  • hen egg lysozyme
  • Muramidase