Increased production of human granulocyte-macrophage colony stimulating factor (hGM-CSF) by the addition of stabilizing polymer in plant suspension cultures

J Biotechnol. 2002 Jul 3;96(3):205-11. doi: 10.1016/s0168-1656(02)00044-5.

Abstract

Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a glycoprotein that stimulates the production of granulocytes, macrophages, and white blood cells. Secretion of human GM-CSF from cell suspension cultures of genetically modified tobacco has been facilitated using natural mammalian leader sequences. At the mid-exponential growth phase (day 4 after the initiation of cell suspension culture), GM-CSF was detected in the medium at a maximum concentration of 180 microg l(-1). However, the secreted GM-CSF was unstable in the medium, and rapidly degraded after day 5. In order to stabilize the secreted GM-CSF, three stabilizing polymers were tested, polyethylene glycol, polyvinylpyrrolidone and gelatin. Gelatin was the most effective in stabilizing the secreted GM-CSF. Following the addition of 5% (w/v) gelatin, the maximum GM-CSF concentration reached 783 microg l(-1), a 4.6-fold increase over control.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Agrobacterium tumefaciens / chemistry
  • Agrobacterium tumefaciens / genetics
  • Cell Division / physiology
  • DNA, Complementary / genetics
  • Endopeptidases / analysis
  • Endopeptidases / metabolism
  • Excipients / metabolism
  • Gelatin / metabolism*
  • Granulocyte-Macrophage Colony-Stimulating Factor / biosynthesis*
  • Granulocyte-Macrophage Colony-Stimulating Factor / genetics
  • Granulocyte-Macrophage Colony-Stimulating Factor / metabolism
  • Humans
  • Nicotiana / cytology*
  • Nicotiana / genetics
  • Nicotiana / metabolism*
  • Nicotiana / microbiology
  • Polyethylene Glycols / metabolism*
  • Povidone / metabolism*
  • Transformation, Bacterial

Substances

  • DNA, Complementary
  • Excipients
  • Polyethylene Glycols
  • Granulocyte-Macrophage Colony-Stimulating Factor
  • Gelatin
  • Endopeptidases
  • Povidone